Anti-Vinculin 抗体 [EPR8185]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Immunofluorescent staining of HEK293 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab129002 at a dilution of 1/50. An Alexa Fluor^® 488 goat anti-rabbit antibody (ab150077) was used as the secondary at a dilution of 1/1000 and the cells were counter stained with DAPI. The negative controls are shown in the bottom middle and right hand panels. For negative control 1, the primary was used and then goat anti-mouse IgG was used at a dilution of 1/500. For negative control 2, a mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used.

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Western blot : Anti-VCL antibody [EPR8185] (ab129002) staining at 1/10000 dilution, shown in green; Mouse anti-CANX [CANX/1543] (ab238078) loading control staining at 1/20000 dilution, shown in magenta. In Western blot, ab129002 was shown to bind specifically to VCL. A band was observed at 124 kDa in wild-type A431 cell lysates with no signal observed at this size in VCL knockout cell line. To generate this image, wild-type and VCL knockout A431 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween^® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.

All lanes:

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (ab129002) at 1/10000 dilution

Lane 1:

Wild-type A431 cell lysate

Lane 2:

Western blot - Human VCL (Vinculin) knockout A-431 cell line (<a href='/products/cell-lines/human-vcl-vinculin-knockout-a-431-cell-line-ab261898'>ab261898</a>) at 20 µg

Lane 2:

VCL knockout A431 cell lysate

Lane 3:

HeLa cell lysate

Lane 4:

Jurkat cell lysate

false

• ICC/IF

Unknown

Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Immunofluorescent staining of vinculin in HUVEC cells with unpurified ab129002 at 1/100 dilution.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Intracellular Flow Cytometry analysis of 293 (human embryonic kidney epithelial) cells labeling Vinculin (red) with ab129002 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr^® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Flow cytometry overlay histogram showing wild-type A-431 (green line) and VCL knockout A-431 stained with ab129002 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab129002) (1x 106 in 100μl at 0.00032 μg/ml (1/6249999)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type A-431 - black line, VCL knockout A-431 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• Flow Cyt (Intra)

Lab

Flow Cytometry (Intracellular) - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Flow cytometry overlay histogram showing left HeLa positive cells and right negative HeLa VCL KO stained with ab129002 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab129002) (1x 10⁶ in 100μl at 0.008μg/ml (1/7875)) for 30min at 22°C.

The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C

Isotype control antibody (black line) was Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.

• IP

Lab

Immunoprecipitation - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

ab129002 (purified) at 1/20 immunoprecipitating vinculin in HeLa cells. Lane 1 : HeLa whole cell lysate (10 μg). Lane 2 : HeLa whole cell lysate (10 μg). Lane 3 : Rabbit monoclonal IgG (ab172730) instead of ab129002 in HeLa whole cell lysate. For western blotting, a HRP-conjugated goat anti-rabbit antibody was used as the secondary antibody (1/1000).

Blocking buffer and concentration : 5% NFDM/TBST.

Diluting buffer and concentration : 5% NFDM /TBST.

All lanes:

Immunoprecipitation - Anti-Vinculin antibody [EPR8185] - Loading Control (ab129002)

Predicted band size: 124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the higher MW band at approximately 300 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-BCL9 antibody [EPR29651-74] (<a href='/products/primary-antibodies/bcl9-antibody-epr29651-74-ab325937'>ab325937</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control fresh whole cell lysate at 20 µg

Lane 2:

293T transfected with siRNA specifically targeting BCL9 fresh whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab325390 was shown to bind specifically to ATRX. Target of interest was observed at 310 kDa in wild-type A549 cell lysates (lane 1) with no signal observed at this size in ATRX knockout cell line (lane 2, knockout cell line ab287232).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-ATRX antibody [EPR29209-280] (<a href='/products/primary-antibodies/atrx-antibody-epr29209-280-ab325390'>ab325390</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 (human lung carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

ATRX knockout A549 whole cell lysate

Lane 3:

Wild-type A549 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 310 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : CT26.WT, U-937.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 20944749, 17925379, 19692571, 25239622).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (124 kDa) staining at 1/10000 dilution.

Exposure time : Lanes 1-2 : 59 seconds, lanes 3-4 : 26 seconds

All lanes:

Western blot - Anti-Pannexin 1 antibody [EPR28631-135] (<a href='/products/primary-antibodies/pannexin-1-antibody-epr28631-135-ab325529'>ab325529</a>) at 1/1000 dilution

Lane 1:

Neuro-2a (mouse neuroblastoma neuroblast) whole cell lysate at 20 µg

Lane 2:

CT26.WT (mouse colon fibroblast) whole cell lysate at 20 µg

Lane 3:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

U-937 (human histiocytic lymphoma monocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 37-50 kDa,124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

293T cells express endogenously BCL9. This antibody does not cross-react with human BCL9L by WB.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa); Anti-6X His tag® antibody [EPR20547] - ChIP Grade (ab213204) (1 : 5000)(200KDa).

All lanes:

Western blot - Anti-BCL9 antibody [EPR29651-74] (<a href='/products/primary-antibodies/bcl9-antibody-epr29651-74-ab325937'>ab325937</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) cells transfected with an empty vector containing a his-tag, fresh whole cell lysate at 20 µg

Lane 2:

293T cells transfected with a human BCL9 expression vector containing a his-tag, fresh whole cell lysate at 20 µg

Lane 3:

293T cells transfected with a human BCL9L expression vector containing a his-tag, fresh whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab325178 was shown to bind specifically to UBP43/USP18. Target of interest was observed at 35 kDa and 37kDa in wide-type A549 cell lysates (lane 2) with no signal observed at this size in USP18 knockout cell line (lane 3, 4) (lane 3, knockout cell line ab267178 / knockout cell lysate ab258752).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-UBP43/USP18 antibody [EPR26174-540] (<a href='/products/primary-antibodies/ubp43-usp18-antibody-epr26174-540-ab325178'>ab325178</a>) at 1/1000 dilution

Lane 1:

Untreated wide-type A549 (human lung carcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

Wide-type A549 treated with 100U/ml IFN alpha (human) for 14 hours whole cell lysate at 50 µg

Lane 3:

Untreated USP18 knockout A549 whole cell lysate at 50 µg

Lane 4:

USP18 knockout A549 treated with 100U/ml IFN alpha (human) for 14 hours whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35 kDa,37 kDa,124 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the higher MW band at approximately 300 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-BCL9 antibody [EPR29651-74] (<a href='/products/primary-antibodies/bcl9-antibody-epr29651-74-ab325937'>ab325937</a>) at 1/1000 dilution

All lanes:

SW620 (human colorectal adenocarcinoma epithelial cell) fresh whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 22558357; PMID : 19166587).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower band at approximately 140 kDa is likely SET-NUP214 fusion protein.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time : Lanes 1-2 : 180 seconds; Lanes 3 : 37 seconds

All lanes:

Western blot - Anti-NUP214 antibody [EPR29100-623] (<a href='/products/primary-antibodies/nup214-antibody-epr29100-623-ab325238'>ab325238</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 268 kDa,124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : HEL, B16-F0, Beta-TC-6.

The identity of the bands lower than 130 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-ITPR3 antibody [EPR25276-304] (<a href='/products/primary-antibodies/itpr3-antibody-epr25276-304-ab325503'>ab325503</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

A-172 (human brain glioblastoma cell ) whole cell lysate at 20 µg

Lane 3:

DLD-1 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

HEL (human erythroleukemia erythroblast) whole cell lysate at 20 µg

Lane 5:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 20 µg

Lane 6:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

B16-F0 (mouse melanoma epithelial-like cell) whole cell lysate at 20 µg

Lane 8:

Beta-TC-6 (mouse pancreas insulinoma beta cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/20000 dilution

Observed band size: 280 kDa,124 kDa

false

Exposure time: 81s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : HEL.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-BCL9 antibody [EPR29651-74] (<a href='/products/primary-antibodies/bcl9-antibody-epr29651-74-ab325937'>ab325937</a>) at 1/1000 dilution

Lane 1:

IMR-32 (human neuroblastoma neuroblast) fresh whole cell lysate at 20 µg

Lane 2:

HEL (human erythroleukemia erythroblast) fresh whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the bands below 300 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

In Western blot, Anti-53BP1 antibody [EPR2172(2)] (ab175933) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-53BP1 (phospho S25) antibody [EPR27057-33] (<a href='/products/primary-antibodies/53bp1-phospho-s25-antibody-epr27057-33-ab315015'>ab315015</a>) at 1/1000 dilution

Lane 1:

Untreated wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell line) whole cell lysate (untreated membrane) at 20 µg

Lane 2:

Wild-type HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (untreated membrane) at 20 µg

Lane 3:

Untreated TP53BP1 knockout HAP1 whole cell lysate (untreated membrane) at 20 µg

Lane 4:

TP53BP1 knockout HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (untreated membrane) at 20 µg

Lane 5:

Untreated wild-type HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 6:

Wild-type HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 7:

Untreated TP53BP1 knockout HAP1 whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Lane 8:

TP53BP1 knockout HAP1 treated with 100nM Calycin A for 30 minutes, whole cell lysate (alkaline phosphatase treated membrane) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 350 kDa,124 kDa

false

Exposure time: 3s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Lanes 1-2 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/100000 and lanes 3-4 are incubated with Goat Anti-Rabbit IgG (HRP) at 1/2000.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time : Lanes 1 and 3-4 : 180 seconds; Lane 2 : 26 seconds

All lanes:

Western blot - Anti-SMC1A antibody [EPR30532-7] (<a href='/products/primary-antibodies/smc1a-antibody-epr30532-7-ab325922'>ab325922</a>) at 1/1000 dilution

Lane 1:

Human spleen tissue lysate at 20 µg

Lane 2:

Human tonsil tissue lysate at 20 µg

Lane 3:

Mouse kidney tissue lysate at 20 µg

Lane 4:

Mouse liver tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Lanes 3 - 4:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 160 kDa,124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab327707 was shown to bind specifically to WRN. Target of interest was observed at 200 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in WRN knockout cell lysates (lane 2).

Low expression : HT-29, PANC-1, SW1116.

The identity of the bands lower than 180 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Werner's syndrome helicase WRN antibody [EPR29645-145] (<a href='/products/primary-antibodies/werners-syndrome-helicase-wrn-antibody-epr29645-145-ab327707'>ab327707</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

WRN knockout HAP1 cell lysate at 20 µg

Lane 3:

K-562 (human chronic myelogenous leukemia lymphoblast) whole cell lysate at 20 µg

Lane 4:

SK-N-MC (human brain epithelial cell) whole cell lysate at 20 µg

Lane 5:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

SW1116 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

Ramos (human burkitt's lymphoma b lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST Low expression : HCT116, HEK-293 (PMID : 23743647). The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 11021599). In lane 1, the lysate was stored at -80°C prior to Western Blotting. The bands beneath the target band (320 kDa) are likey to be degradation products. In lanes 2-4, the lysates were freshly made and used for Western Blotting immediately to minimize protein degradation. In Western blot, anti- Vinculin antibody (ab129002) loading control staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Collagen VII antibody [EPR27012-157] (<a href='/products/primary-antibodies/collagen-vii-antibody-epr27012-157-ab309143'>ab309143</a>) at 1/1000 dilution

Lane 1:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HCT116 (human colorectal carcinoma epithelial cell), whole cell lysate at 20 µg

Lane 4:

HEK-293 (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 320 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : brain.

The molecular weight is consistent with what have been described in literature (PMID : 12582180; 16754877).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Exposure time : Lanes 1, 3 and 4 : 180 seconds; Lane 2 : 37 seconds

All lanes:

Western blot - Anti-MUC2 antibody [RM1281] (<a href='/products/primary-antibodies/muc2-antibody-rm1281-ab322964'>ab322964</a>) at 1/1000 dilution

Lane 1:

HT-29 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

Mouse colon tissue lysate at 20 µg

Lane 3:

Mouse small intestine tissue lysate at 20 µg

Lane 4:

Mouse brain tissue lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 140-170 kDa,124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : PANC-1, G-401, skeletal muscle.

The identity of the lower MW band at approximately 130 kDa (in lane 4) is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-KIF13B antibody [EPR29559-519] (<a href='/products/primary-antibodies/kif13b-antibody-epr29559-519-ab327425'>ab327425</a>) at 1/1000 dilution

Lane 1:

NCI-H929 (human Bone B lymphocyte) whole cell lysate at 60 µg

Lane 2:

PANC-1 (human pancreatic epithelioid carcinoma epithelial cell) whole cell lysate at 60 µg

Lane 3:

G-401 (human rhabdoid tumor kidney epithelial cell) whole cell lysate at 60 µg

Lane 4:

Mouse liver tissue lysate at 60 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 60 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 260 kDa,124 kDa

false

Exposure time: 158s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab324875 was shown to bind specifically to Tet2 . Target of interest was observed at 260 kDa in wild-type HAP1 cell lysates (lane 1) with no signal observed at this size in Tet2 knockout cell line (lane 2).

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The bands beneath the target band (260 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Tet2 antibody [EPR29630-29] (<a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a>) at 1/1000 dilution

Lane 1:

Wild-type HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

Tet2 knockout HAP1 whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 260 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-KIF13B antibody [EPR29559-519] (<a href='/products/primary-antibodies/kif13b-antibody-epr29559-519-ab327425'>ab327425</a>) at 1/1000 dilution

Lane 1:

A-204 (human muscle rhabdomyosarcoma cell) transfected with scrambled siRNA control whole cell lysate at 80 µg

Lane 2:

A-204 transfected with siRNA specifically targeting KIF13B whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - VeriBlot for IP Detection Reagent (HRP) (<a href='/products/reagents/veriblot-for-ip-detection-reagent-hrp-ab131366'>ab131366</a>) at 1/1000 dilution

Observed band size: 260 kDa,124 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The bands beneath the target band (260 kDa) are likely to be degraded target fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-Tet2 antibody [EPR29630-29] (<a href='/products/primary-antibodies/tet2-antibody-epr29630-29-ab324875'>ab324875</a>) at 1/1000 dilution

Lane 1:

293T (human embryonic kidney epithelial cell) transfected with scrambled siRNA control whole cell lysate at 10 µg

Lane 2:

293T transfected with siRNA specifically targeting Tet2 whole cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 260 kDa,124 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Low expression : heart (PMID : 23591958).

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

Samples should not be boiled, as boiling may cause protein aggregation.

All lanes:

Western blot - Anti-CerS2 antibody [EPR28098-1] (<a href='/products/primary-antibodies/cers2-antibody-epr28098-1-ab315452'>ab315452</a>) at 1/1000 dilution

Lane 1:

HepG2 (human hepatocellar carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Mouse heart tissue lysate at 20 µg

Lane 4:

Rat brain tissue lysate at 20 µg

Lane 5:

Rat heart tissue lysate at 20 µg

Lane 6:

Hepa1-6 (mouse hepatoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 40 kDa,124 kDa

false

Exposure time: 15s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

This blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

To minimize protein degradation, cells were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

In Western blot, Anti-Vinculin antibody [EPR8185] ab129002 staining at 1/10000 dilution.

All lanes:

Western blot - Anti-KDM6B / JMJD3 antibody [EPR28325-184] (<a href='/products/primary-antibodies/kdm6b-jmjd3-antibody-epr28325-184-ab324740'>ab324740</a>) at 1/1000 dilution

Lane 1:

K-562 (human chronic myelogenous leukemia lymphoblast) transfected with scrambled siRNA control whole cell fresh lysate at 20 µg

Lane 2:

K-562 transfected with siRNA specifically targeting KDM6B whole cell fresh lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 200 kDa,124 kDa

true

Exposure time: 92s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The identity of the bands lower than 180 kDa are unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Werner's syndrome helicase WRN antibody [EPR29645-145] (<a href='/products/primary-antibodies/werners-syndrome-helicase-wrn-antibody-epr29645-145-ab327707'>ab327707</a>) at 1/1000 dilution

All lanes:

Human testis tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, ab327426 was shown to bind specifically to CCNE1. Target of interest was observed at 35-50 kD in wild-type HCT116 cell lysates (lanes 1-2) with no signal observed at this size in the CCNE1 knockout cell line (lanes 3-4) (lanes 3-4, knockout cell line ab286494).

The identity of the higher MW band at approximately 150 kDa is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Cyclin E1(Phospho T77) antibody [EPR28889-10] (<a href='/products/primary-antibodies/cyclin-e1phospho-t77-antibody-epr28889-10-ab327426'>ab327426</a>) at 1/1000 dilution

Lane 1:

Untreated HCT 116 (human colorectal carcinoma epithelial cell) whole cell lysate (untreated membrane) at 50 µg

Lane 2:

HCT 116 treated with 2.5mM Thymidine for 16h whole cell lysate (untreated membrane) at 50 µg

Lane 3:

CCNE1 knockout HCT116 whole cell lysate (untreated membrane) at 50 µg

Lane 4:

CCNE1 knockout HCT116 treated with 2.5mM Thymidine for 16h) whole cell lysate (untreated membrane) at 50 µg

Lane 5:

Untreated HCT 116 whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 6:

HCT 116 treated with 2.5mM Thymidine for 16h whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 7:

CCNE1 knockout HCT116 whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Lane 8:

CCNE1 knockout HCT116 treated with 2.5mM Thymidine for 16h whole cell lysate (alkaline phosphatase treated membrane) at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 35-50 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Performed under reducing conditions.

In Western blot, ab327421 was shown to bind specifically to Talin 1. Target of interest was observed at 269, 250, 240 kDa in wild-type PC-3 cell lysates (lane 1) with no signal observed at this size in TLN1 knockout cell line (lane 2) (lane 2, knockout cell line ab290495).

Low expression : TT.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Talin 1 antibody [EPR29641-30] (<a href='/products/primary-antibodies/talin-1-antibody-epr29641-30-ab327421'>ab327421</a>) at 1/1000 dilution

Lane 1:

Wild-type PC-3 (human prostate adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Western blot - Human TLN1 knockout PC-3 cell line (<a href='/products/cell-lines/human-tln1-knockout-pc-3-cell-line-ab290495'>ab290495</a>) at 20 µg

Lane 3:

NIH/3T3 (mouse embryonic fibroblast) whole cell lysate at 20 µg

Lane 4:

bEnd.3 (mouse brain endothelial cell) whole cell lysate at 20 µg

Lane 5:

SK-OV-3 (human ovarian cancer epithelial cell) whole cell lysate at 20 µg

Lane 6:

LN-229 (human brain glioblastoma epithelial cell) whole cell lysate at 20 µg

Lane 7:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

TT (human thyroid carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 269 kDa,250 kDa,240 kDa,124 kDa

false

Exposure time: 70s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, the fresh lysates were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW bands are likely to be degradation fragments.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-ASXL2 antibody [EPR27482-289] (<a href='/products/primary-antibodies/asxl2-antibody-epr27482-289-ab327007'>ab327007</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

293T (human embryonic kidney epithelial cell) whole cell lysate at 20 µg

Lane 3:

THP-1 (human monocytic leukemia monocyte) whole cell lysate at 20 µg

Lane 4:

Jurkat (human T cell leukemia T lymphocyte from peripheral blood) whole cell lysate at 20 µg

Lane 5:

RAW 264.7 (mouse Abelson murine leukemia virus-induced tumor macrophage) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 210 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

To minimize protein degradation, the fresh lysates were lysed immediately after harvest and then applied to a gel and transfer membrane for Western blotting as soon as possible.

The identity of the lower MW bands are likely to be degradation fragments.

Lanes 3-4 of this blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-ASXL2 antibody [EPR27482-289] (<a href='/products/primary-antibodies/asxl2-antibody-epr27482-289-ab327007'>ab327007</a>) at 1/1000 dilution

Lane 1:

3T3-L1 (mouse embryonic fibroblast) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

3T3-L1 transfected with siRNA specifically targeting ASXL2 whole cell lysate at 20 µg

Lane 3:

U-2 OS (human bone osteosarcoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 4:

U-2 OS transfected with siRNA specifically targeting ASXL2 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 210 kDa,124 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : spleen, kidney, cerebellum.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 36054350).

Samples are non-boiled as boiling may cause protein aggregation.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time : Lanes 1-4 : 59 seconds, lanes 5-7 : 180 seconds

All lanes:

Western blot - Anti-Muscarinic Acetylcholine Receptor 1/CHRM1 antibody [EPR26494-240] (<a href='/products/primary-antibodies/muscarinic-acetylcholine-receptor-1-chrm1-antibody-epr26494-240-ab325432'>ab325432</a>) at 1/1000 dilution

Lane 1:

Mouse cerebral cortex tissue lysate at 50 µg

Lane 2:

Mouse kidney tissue lysate at 50 µg

Lane 3:

Mouse spleen tissue lysate at 50 µg

Lane 4:

Mouse cerebellum tissue lysate at 50 µg

Lane 5:

Human cerebral cortex tissue lysate at 50 µg

Lane 6:

Human spleen tissue lysate at 50 µg

Lane 7:

Human cerebellum tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 75 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-UBP43/USP18 antibody [EPR26174-540] (<a href='/products/primary-antibodies/ubp43-usp18-antibody-epr26174-540-ab325178'>ab325178</a>) at 1/1000 dilution

All lanes:

Human tonsil tissue lysate at 50 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 35 kDa,37 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

The expression of NLRC5 is upregulated in response to IFN gamma treatment (PMID : 36103539; PMID : 20610642; PMID : 20061403).

This antibody can recognize the different isoforms of NLRC5.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-NLRC5 antibody [EPR29722-55] (<a href='/products/primary-antibodies/nlrc5-antibody-epr29722-55-ab325273'>ab325273</a>) at 1/1000 dilution

Lane 1:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

HeLa treated with 1000U/ml IFN gamma for 6 hours whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 205 kDa,170 kDa,124 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : human liver, human spleen.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time :
Lanes 1-3 : 10 seconds;
Lane 4 : 48 seconds.

All lanes:

Western blot - Anti-EMP2 antibody [EPR30305-511] (<a href='/products/primary-antibodies/emp2-antibody-epr30305-511-ab326423'>ab326423</a>) at 1/1000 dilution

Lane 1:

Human lung tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human spleen tissue lysate at 20 µg

Lane 4:

A431 (human epidermoid carcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 20-50 kDa,124 kDa

false

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : 293T(PMID : 10934049)

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) 1 : 10000 (124Kda).

All lanes:

Western blot - Anti-PTPN14 antibody [EPR30487-534] (<a href='/products/primary-antibodies/ptpn14-antibody-epr30487-534-ab326098'>ab326098</a>) at 1/1000 dilution

Lane 1:

DU 145 (human prostate carcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 80 µg

Lane 2:

DU 145 transfected with siRNA specifically targeting PTPN14 whole cell lysate at 80 µg

Lane 3:

293T (human embryonic kidney epithelial cell) whole cell lysate at 80 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 135 kDa,124 kDa

false

Exposure time: 135s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : lung.

The molecular weight observed is consistent with what has been described in the literature (PMID : 18940796).

The identity of the higher MW band at approximately 250 kDa (in lane 1) is unknown.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-ITGA7 antibody [EPR28509-556] (<a href='/products/primary-antibodies/itga7-antibody-epr28509-556-ab325935'>ab325935</a>) at 1/1000 dilution

Lane 1:

Human skeletal muscle tissue lysate at 40 µg

Lane 2:

Human bladder tissue lysate at 40 µg

Lane 3:

Human lung tissue lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 70 kDa,100 kDa,128 kDa,124 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-UBP43/USP18 antibody [EPR26174-540] (<a href='/products/primary-antibodies/ubp43-usp18-antibody-epr26174-540-ab325178'>ab325178</a>) at 1/1000 dilution

Lane 1:

Untreated THP-1 (human monocytic leukemia monocyte) whole cell lysate at 50 µg

Lane 2:

THP-1 treated with 1 ug/ml LPS for 8 hours whole cell lysate at 50 µg

Lane 3:

Untreated HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

HeLa treated with 10ng/ml IFN alpha (human) for 16 hours whole cell lysate at 50 µg

Lane 5:

Untreated MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 6:

MCF7 treated with 100U/ml IFN alpha (human) for 24 hours whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/100000 dilution

Observed band size: 35 kDa,37 kDa,124 kDa

false

Exposure time: 6s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : spleen.

250 kDa full length band and 120-130 kDa cleavage fragment band were ovserved (PMID : 20471383).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Robo1 antibody [EPR29534-534] (<a href='/products/primary-antibodies/robo1-antibody-epr29534-534-ab326048'>ab326048</a>) at 1/1000 dilution

Lane 1:

Human breast cancer tissue lysate at 20 µg

Lane 2:

Human liver cancer tissue lysate at 20 µg

Lane 3:

Human colon cancer tissue lysate at 20 µg

Lane 4:

Human spleen tissue lysate at 20 µg

Secondary

All lanes:

Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 dilution

Observed band size: 120-130 kDa,124 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

Low expression : MCF7, LoVo.

250 kDa full length band and 120-130 kDa cleavage fragment band were ovserved (PMID : 20471383).

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

All lanes:

Western blot - Anti-Robo1 antibody [EPR29534-534] (<a href='/products/primary-antibodies/robo1-antibody-epr29534-534-ab326048'>ab326048</a>) at 1/1000 dilution

Lane 1:

HAP1 (human chronic myelogenous leukemia near-haploid cell) whole cell lysate at 20 µg

Lane 2:

U-87 MG (human glioblastoma-astrocytoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 4:

MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 6:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 120-130 kDa,250 kDa,124 kDa

false

Exposure time: 37s

• WB

Lab

Western blot - Anti-Vinculin antibody [EPR8185] - Loading Control (AB129002)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.

In Western blot, Anti-Vinculin antibody [EPR8185] (ab129002) (1 : 10000) (124KDa).

Exposure time : Lanes 1-3 : 10 seconds, lanes 4-5 : 180 seconds

All lanes:

Western blot - Anti-INO80 antibody [EPR30384-761] (<a href='/products/primary-antibodies/ino80-antibody-epr30384-761-ab325542'>ab325542</a>) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 2:

LoVo (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 3:

SW480 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 50 µg

Lane 4:

CT26.WT (mouse colon fibroblast) whole cell lysate at 50 µg

Lane 5:

B16-F10 (mouse skin melanoma cell) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Observed band size: 200 kDa,124 kDa

false