Goat Anti-Rabbit IgG H&L (HRP)

41 Images

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-RAB29 (phospho T71) antibody [MJF-R24-17-1] (<a href='/products/primary-antibodies/rab29-phospho-t71-antibody-mjf-r24-17-1-ab241062'>ab241062</a>) at 1/5000 dilution

Lane 1:

HEK-293 (human embryonic kidney) over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

Lane 2:

MLi-2 (100nM for 60min) treated HEK-293 over-expressing HA-tagged RAB29 and LRRK2 [Y1699C], whole cell lysate at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 23 kDa

Observed band size: 23 kDa

false

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Immunohistochemical analysis of paraffin-embedded Human skin tissue labeling Histone H3 (mono methyl K79) with ab177183 at 1/5000 dilution, followed by ab97051 Goat Anti-Rabbit HRP (IgG H&L) at 1/500 dilution. Nucleus staining on Human skin tissue is observed. Counter stained with Hematoxylin.

Negative control : Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

• IHC-P

Lab

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

ab92511 staining HLA-DR in human spleen tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and antigen retrieval was by heat mediation in a EDTA buffer. Samples were incubated with primary antibody at a dilution of 1/700. A goat anti-rabbit IgG H&L (HRP) ab97051 was used as the secondary antibody at a dilution of 1/500. Negative control 1 : PBS in place of primary antibody.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

• WB

Unknown

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 24517394).

Negative control : Human liver (PMID : 28611447).

HepG2 as a low serpinB expressing cell line.

This blot was developed using a higher sensitivity ECL substrate.

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-SerpinB3/SCCA + SerpinB4/SCCA-2 antibody [EPR22455-57] (<a href='/products/primary-antibodies/serpinb3-scca-serpinb4-scca-2-antibody-epr22455-57-ab254255'>ab254255</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

Human fetal lung tissue lysate at 20 µg

Lane 4:

Human liver tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 45 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

All lanes:

Western blot - Anti-Neurogenin 2/NGN2 antibody [EPR4177] (<a href='/products/primary-antibodies/neurogenin-2-ngn2-antibody-epr4177-ab109236'>ab109236</a>) at 1/5000 dilution

All lanes:

SH-SY5Y cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 28 kDa

false

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and dilution buffer : 5% NFDM /TBST.

All lanes:

Western blot - Anti-Ionotropic glutamate receptor 2 + 3 antibody [EP929Y] (<a href='/products/primary-antibodies/ionotropic-glutamate-receptor-2-3-antibody-ep929y-ab52896'>ab52896</a>) at 1/2000 dilution

All lanes:

Human fetal brain tissue lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 98 kDa

Observed band size: 100 kDa

false

• WB

Unknown

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

All lanes:

Western blot - Anti-SNF2L antibody (<a href='/products/primary-antibodies/snf2l-antibody-ab37003'>ab37003</a>) at 1 µg/mL

Lane 1:

Jurkat nuclear extract lysate (<a href='/products/unavailable/jurkat-nuclear-extract-lysate-ab14844'>ab14844</a>) at 20 µg

Lane 2:

HEK293 (Human embryonic kidney cell line) Nuclear Lysate at 20 µg

Lane 3:

SHSY5Y Whole Cell Lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 123 kDa

Observed band size: 140 kDa,70 kDa

true

Exposure time: 16min

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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 : 28541286 and PMID : 157438355).

Exposure time : Lane 1 : 180 seconds; Lanes 2-7 : 59 seconds

All lanes:

Western blot - Anti-PSAP antibody [RM1353] (ab324885) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Lane 4:

Mouse cerebellum tissue lysate at 20 µg

Lane 5:

Mouse testis tissue lysate at 20 µg

Lane 6:

Rat testis tissue lysate at 20 µg

Lane 7:

Rat cerebellum tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 60-80 kDa,12 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking/Dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-Histone H2B (formyl K108) antibody [EPR17697] (<a href='/products/primary-antibodies/histone-h2b-formyl-k108-antibody-epr17697-ab177784'>ab177784</a>) at 1/2000 dilution

All lanes:

Hela (Human epithelial cells from cervix adenocarcinoma) cell lysate at 10 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 14 kDa

Observed band size: 14 kDa

false

Exposure time: 10s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

This blot was produced using a 4-12% Bis-tris gel under the MES buffer system. The gel was run at 200V for 35 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab125712 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

All lanes:

Western blot - Anti-MOX1/MEOX1 antibody (<a href='/products/primary-antibodies/mox1-meox1-antibody-ab125712'>ab125712</a>) at 1 µg/mL

All lanes:

Heart (Human) Whole Cell Lysate - fetal normal tissue at 25 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

Predicted band size: 27 kDa

Observed band size: 34 kDa,65 kDa,75 kDa

true

Exposure time: 20min

• WB

Unknown

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

This blot was produced using a 10% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% milk before being incubated with ab10134 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution. The band detected by ab10134 migrates at approximately 60-kDa. Although the predicted molecular weight of this protein is 72-kDa, the 60-kDa band has been observed in the literature. The intensity of this band has been shown to increase in cells exposed to hypoxic conditions (PMID : 16775626).

Abcam welcomes customer feedback and would appreciate any comments regarding this product and the data presented above.

All lanes:

Anti-HIF3 alpha/IPAS antibody (ab10134) at 2 µg/mL

All lanes:

Human heart tissue lysate - total protein (<a href='/products/unavailable/human-heart-tissue-lysate-total-protein-ab29431'>ab29431</a>) at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

true

Exposure time: 8min

• WB

Unknown

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

All lanes:

Western blot - Anti-E Cadherin antibody [EPR699] (<a href='/products/primary-antibodies/e-cadherin-antibody-epr699-ab133597'>ab133597</a>) at 1/1000 dilution

All lanes:

MCF7 (Human breast adenocarcinoma epithelial cell) whole cell lysates at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 97 kDa

Observed band size: 120 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : liver

Lanes 1-2 are incubated with are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 3-11 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PFKP antibody [EPR28830-157] (ab325016) at 1/1000 dilution

Lane 1:

Human colon cancer tissue lysate at 20 µg

Lane 2:

Human lung cancer tissue lysate at 20 µg

Lane 3:

Mouse testis tissue lysate at 20 µg

Lane 4:

Rat testis tissue lysate at 20 µg

Lane 5:

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

Lane 6:

P815 (mouse mastocytoma mast Cell ) whole cell lysate at 20 µg

Lane 7:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 8:

Mouse brain tissue lysate at 20 µg

Lane 9:

Mouse liver tissue lysate at 20 µg

Lane 10:

Rat brain tissue lysate at 20 µg

Lane 11:

Rat liver tissue lysate at 20 µg

Secondary

Lanes 1 - 2:

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

Lanes 3 - 11:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 85 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and dilution buffer : 5% NFDM/TBST.

The molecular mass observed is consistent with what has been described in the literature (PMID : 26482643).

All lanes:

Western blot - Anti-DDX4 / MVH antibody [EPR21785] (<a href='/products/primary-antibodies/ddx4-mvh-antibody-epr21785-ab235443'>ab235443</a>) at 1/1000 dilution

All lanes:

Human testis lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 79 kDa

Observed band size: 79 kDa

false

Exposure time: 6s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Lysates were freshly made and used for Western blotting immediately to minimize protein degradation.

All lanes:

Western blot - Anti-KMT6 / EZH2 antibody [EPR25353-284] (<a href='/products/primary-antibodies/kmt6-ezh2-antibody-epr25353-284-ab307646'>ab307646</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Lane 4:

PC-12 (rat adrenal gland pheochromocytoma) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 85 kDa

false

Exposure time: 26s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : skeletal muscle.

Lanes 1-7 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000 and lanes 8-9 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-4 : 1 second, lanes 5-9 : 48 seconds

All lanes:

Western blot - Anti-Snapin antibody [RM2091] (<a href='/products/primary-antibodies/snapin-antibody-rm2091-ab324947'>ab324947</a>) at 1/1000 dilution

Lane 1:

Mouse brain tissue lysate at 50 µg

Lane 2:

Mouse testis tissue lysate at 50 µg

Lane 3:

Mouse spleen tissue lysate at 50 µg

Lane 4:

Mouse skeletal muscle tissue lysate at 50 µg

Lane 5:

Rat brain tissue lysate at 50 µg

Lane 6:

Rat testis tissue lysate at 50 µg

Lane 6:

Rat skeletal muscle tissue lysate at 50 µg

Lane 8:

Human cerebellum tissue lysate at 50 µg

Lane 9:

Human testis tissue lysate at 50 µg

Secondary

Lanes 1 - 7:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Lanes 8 - 9:

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

Observed band size: 15 kDa,36 kDa

false

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

ab40776 may detect non-specific signals due to sample batch variance.

All lanes:

Western blot - Anti-PI 3 Kinase catalytic subunit alpha/PIK3CA antibody [EP383Y] (<a href='/products/primary-antibodies/pi-3-kinase-catalytic-subunit-alpha-pik3ca-antibody-ep383y-ab40776'>ab40776</a>) at 1/1000 dilution

Lane 1:

Batch 1 MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 2:

Batch 2 MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

Batch 3 MCF7 (human breast adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 110 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-EGFR antibody - Total protein control (ab32077) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-EGFR (phospho Y1045) antibody [EPR28381-88] (<a href='/products/primary-antibodies/egfr-phospho-y1045-antibody-epr28381-88-ab316155'>ab316155</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

A431 treated with 100 ng/ml EGF for 30 minutes whole cell lysate (untreated membrane) at 20 µg

Lane 3:

A431 treated with 100 ng/ml EGF 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) (ab97051) at 1/100000 dilution

Observed band size: 175 kDa,36 kDa

false

Exposure time: 10s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and diluting buffer and concentration : 5% NFDM/TBST. Negative control : Raji (PMID : 8144934, PMID : 10583368, PMID : 1966549). The molecular weight observed is consistent with the isoforms. In Western blot, anti-GAPDH antibody (ab181602) loading control staining at 1/200,000 dilution. Exposure time : Lane 1 : 92 seconds, Lanes 2-4 : 180 seconds.

All lanes:

Western blot - Anti-TNF Receptor I antibody [EPR26198-14] (<a href='/products/primary-antibodies/soluble-tnf-receptor-i-fc-fragment-antibody-epr26198-14-ab308506'>ab308506</a>) at 1/1000 dilution

Lane 1:

A549 (human lung carcinoma epithelial cell) whole cell lysate, at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

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

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Predicted band size: 50 kDa

Observed band size: 55 kDa,38 kDa,28 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : NCI-H1299, Calu-1 (PMID : 35395804)

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 35395804).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Ninjurin-1 antibody [EPR30446-757] (ab325137) at 1/1000 dilution

Lane 1:

NCI-H1975 (human non-small cell lung cancer epithelial cell), whole cell lysate at 20 µg

Lane 3:

A549 (human lung carcinoma epithelial cell), whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 16 kDa,36 kDa

false

Exposure time: 114s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : RAW 264.7, Beta-TC-6, Raji, HDLM-2

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 19936252; PMID : 26847765; PMID : 17119023).

Lanes 1-4 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) staining at 1/10000 dilution.

All lanes:

Western blot - Anti-LRP4 antibody [EPR30692-584] (ab324976) at 1/1000 dilution

Lane 1:

F9 (mouse embryonal carcinoma epithelial cell) whole cell lysate at 48 µg

Lane 2:

MEF (mouse embryo fibroblast) whole cell lysate at 48 µg

Lane 5:

SK-N-BE(2) (human neuroblastoma neuroblast) whole cell lysate at 48 µg

Lane 7:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 48 µg

Lane 8:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 48 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 270 kDa,124 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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 bands lower than 25 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PALB2 antibody [EPR26990-22] (ab325017) 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:

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

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 131 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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 identity of the bands lower than 75 kDa are unknown.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PALB2 antibody [EPR26990-22] (ab325017) at 1/1000 dilution

Lane 1:

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

Lane 2:

293T transfected with siRNA specifically targeting PALB2 whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 131 kDa,36 kDa

true

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Lanes 1 - 6:

Western blot - Anti-Met (c-Met) antibody [EP1454Y] - N-terminal (<a href='/products/primary-antibodies/met-c-met-antibody-ep1454y-n-terminal-ab51067'>ab51067</a>) at 1/1000 dilution

Lanes 1 - 2:

Western blot - Anti-IMPDH2 antibody [EPR8365(B)] (<a href='/products/primary-antibodies/impdh2-antibody-epr8365b-ab129165'>ab129165</a>) at 1/1000 dilution

Lane 1:

Wild-type A549 cell lysate at 20 µg

Lane 2:

MET knockout A549 cell lysate at 20 µg

Lane 3:

Wild-type HeLa ab255929 cell lysate at 20 µg

Lane 4:

MET (Met (c-Met)) knockout HeLa <a href='/products/cell-lysates/human-met-c-met-knockout-hela-cell-lysate-ab256991'>ab256991</a> cell lysate at 20 µg

Lane 5:

HT-29 cell lysate at 20 µg

Lane 6:

T-47D cell lysate at 20 µg

Secondary

All lanes:

NFDM/TBST at 1/20000 dilution

Observed band size: 40-50 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : MeWo.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lane 1 : 26 seconds; Lanes 2-3 : 81 seconds.

All lanes:

Western blot - Anti-LPCAT3 antibody [EPR28798-172] (ab324941) at 1/1000 dilution

Lane 1:

KARPAS-299 (human t cell lymphoma cell) whole cell lysate at 20 µg

Lane 2:

MeWo (human malignant melanoma fibroblast) whole cell lysate at 20 µg

Lane 3:

RPMI-8226 (human plasmacytoma b lymphocyte) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 40 kDa,36 kDa

false

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

This blot was produced using a 3-8% Tris Acetate gel under the TA buffer system. The gel was run at 150V for 60 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Bovine Serum Albumin before being incubated with ab129945 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution.

All lanes:

Western blot - Anti-C3 antibody (<a href='/products/primary-antibodies/c3-antibody-ab129945'>ab129945</a>) at 1 µg/mL

Lanes 1 - 2:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 3:

HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate at 20 µg with Immunising peptide

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/10000 dilution

Predicted band size: 187 kDa

Observed band size: 200 kDa,50 kDa,55 kDa

true

Exposure time: 8min

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : SH-SY5Y.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

Exposure time : Lanes 1-2 : 70 seconds; Lanes 3-4 : 8 seconds.

All lanes:

Western blot - Anti-LPCAT3 antibody [EPR28798-172] (ab324941) at 1/1000 dilution

Lane 1:

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

Lane 2:

HEK-293 transfected with siRNA specifically targeting LPCAT3 whole cell lysate at 20 µg

Lane 3:

HT-1197 (human urinary bladder carcinoma cell ) whole cell lysate at 20 µg

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 40 kDa,36 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : U-937.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Sp8 antibody [EPR29574-561] (ab324637) at 1/1000 dilution

All lanes:

NCI-H69 (human lung small cell carcinoma cell line) whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 50 kDa,36 kDa

false

Exposure time: 180s

• WB

Lab

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

Blocking and diluting buffer and concentration : 5% NFDM/TBST.
ab181602 was used as a loading control at a 1/200000 dilution.

The expression of PINK1 is upregulated in response to valinomycin treatment (PMID : 22724072).

Left panel is incubated with ab186303 followed by secondary antibody Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/2000 dilution and right panel is incubated with ab323807 followed by Peroxidase-Conjugated Goat anti-Mouse IgG (H+L) at 1/20000 dilution.

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

The blot of right panel was developed using a ECL substrate, allowing for the detection of proteins in the picogram to high-femtogram range.

All lanes:

Western blot - Anti-PINK1 antibody [N4/15] (<a href='/products/primary-antibodies/pink1-antibody-n4-15-ab186303'>ab186303</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SK-OV-3 treated with 30μM CCCP for 6h, whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

Predicted band size: 62 kDa

true

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : liver, skeletal muscle (PMID : 10820318).

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34149921, PMID : 10820318).

The identity of the band at approximately 37 kDa in lanes 5 is unknown.

Lanes 1-3 are incubated with Goat Anti-Rabbit IgG (HRP) with minimal cross-reactivity with human IgG at 1/2000 and lanes 4-5 are incubated with Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated (ab97051) at 1/20000.

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-MIST-1 antibody [EPR29652-63] (ab324434) at 1/1000 dilution

Lane 1:

Human pancreas tissue lysate at 20 µg

Lane 2:

Human liver tissue lysate at 20 µg

Lane 3:

Human skeletal muscle tissue lysate at 20 µg

Lane 4:

Mouse pancreas tissue lysate at 20 µg

Lane 5:

Mouse skeletal muscle tissue lysate at 20 µg

Secondary

Lanes 1 - 3:

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

Lanes 4 - 5:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 21 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Low expression : KATO-III (PMID : 34149921), HepG2, SK-BR-3.

Lanes 1-2 of this blot was developed using a high-sensitivity ECL substrate, allowing for the detection of proteins in the mid-femtogram range.

The identity of the bands higher than 25 kDa in lanes 1-2 are unknown.

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 34149921, PMID : 10820318).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-MIST-1 antibody [EPR29652-63] (ab324434) at 1/1000 dilution

Lane 1:

SW620 (human colorectal adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Lane 2:

KATO III (human stomach spherical) whole cell lysate at 40 µg

Lane 3:

LNCaP (human prostate carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 4:

HepG2 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 40 µg

Lane 5:

SK-BR-3 (human breast adenocarcinoma epithelial cell) whole cell lysate at 40 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 21 kDa,36 kDa

true

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Ret antibody (ab134100) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Ret (phospho Y1015) antibody [EPR26919-93] (ab324978) at 1/1000 dilution

Lanes 1 and 3:

Untreated TT (human thyroid carcinoma epithelial cell) whole cell lysate at 50 µg

Lanes 2 and 4:

TT treated with 100uM pervanadate for 10 min whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PFKP antibody [EPR28830-157] (ab325016) at 1/1000 dilution

Lane 1:

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

Lane 2:

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

Lane 3:

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

Lane 4:

DU 145 (human prostate carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

Raji (human Burkitt's lymphoma B lymphocyte) whole cell lysate at 20 µg

Lane 6:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 85 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

In Western blot, ab324947 was shown to bind specifically to Snapin. Target of interest was observed at 15 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in SNAPIN knockout cell line (lane 2).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-Snapin antibody [RM2091] (<a href='/products/primary-antibodies/snapin-antibody-rm2091-ab324947'>ab324947</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

SNAPIN knockout HeLa whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 15 kDa,36 kDa

false

Exposure time: 37s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Performed under reducing conditions.

In Western blot, ab325016 was shown to bind specifically to PFKP. Target of interest was observed at 85 kDa in wild-type HeLa cell lysates (lane 1) with no signal observed at this size in PFKP knockout cell line (lane 2) (lane 2, knockout cell line ab265136/ knockout cell lysate ab257580).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

All lanes:

Western blot - Anti-PFKP antibody [EPR28830-157] (ab325016) at 1/1000 dilution

Lane 1:

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

Lane 2:

PFKP knockout HeLa whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 85 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

Negative control : T-84, JAR

In lanes 1-4, 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 lanes 5-8, the lysates were stored at -80°C prior to Western Blotting.

The bands beneath the target bands(327 kDa) are expected to be degradation products.

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

Exposure time : Lanes 1-2 : 158 seconds; Lanes 3-8 : 180 seconds

All lanes:

Western blot - Anti-Neurobeachin antibody [EPR30440-555] (<a href='/products/primary-antibodies/neurobeachin-antibody-epr30440-555-ab324953'>ab324953</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

T84 (human colon epithelial cell) fresh whole cell lysate at 20 µg

Lane 3:

Y79 (human retinoblastoma cell) fresh whole cell lysate at 20 µg

Lane 4:

JAR (human placenta choriocarcinoma epithelial cell) fresh whole cell lysate at 20 µg

Lane 5:

Y79 (human retinoblastoma cell) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

T84 (human colon epithelial cell) whole cell lysate at 20 µg

Lane 8:

JAR (human placenta choriocarcinoma epithelial cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 327 kDa,130-327 kDa,124 kDa

false

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

All lanes:

Western blot - Anti-Ninjurin-1 antibody [EPR30446-757] (ab325137) at 1/1000 dilution

Lane 1:

Human colon tissue lysate at 20 µg

Lane 2:

Human lung tissue lysate at 20 µg

Lane 3:

Mouse spleen tissue lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 16 kDa

false

Exposure time: 114s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

In Western blot, ab316155 was shown to bind specifically to EGFR (phospho Y1045). Target of interest was observed at 175 kDa in wild-type HeLa cell lysates (lane 2) with no signal observed at this size in EGFR knockout cell line (lanes 3-4 KO cell line ab255385/KO cell lysate ab263845).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-EGFR antibody (ab32077) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-EGFR (phospho Y1045) antibody [EPR28381-88] (<a href='/products/primary-antibodies/egfr-phospho-y1045-antibody-epr28381-88-ab316155'>ab316155</a>) at 1/1000 dilution

Lane 1:

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

Lane 2:

Wild-type HeLa treated with 100 ng/ml EGF for 30 minutes whole cell lysate at 20 µg

Lane 3:

Untreated EGFR knockout HeLa whole cell lysate at 20 µg

Lane 4:

EGFR knockout HeLa treated with 100 ng/ml EGF for 30 minutes whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 175 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

LC-2/ad cells express the constitutively active CCDC6-RET fusion. The down-regulation of Ret phospho Y1015 is induced by ponatinib treatment (PMID : 33795352) .

The molecular weight observed is consistent with what has been described in the literature (PMID : 33795352; PMID : 23578175).

In Western blot, Anti-GAPDH antibody [EPR16891] - Loading Control (ab181602) staining at 1/200000 dilution.

In Western blot, Anti-Ret antibody (ab134100) staining at 1/1000 dilution.

All lanes:

Western blot - Anti-Ret (phospho Y1015) antibody [EPR26919-93] (ab324978) at 1/1000 dilution

Lanes 1 and 3:

Untreated LC-2/ad (human Lung Epithelial) whole cell lysate at 50 µg

Lanes 2 and 4:

LC-2/ad treated with 300nM Ponatinib for 2h whole cell lysate at 50 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

Observed band size: 60 kDa,36 kDa

false

Exposure time: 180s

• WB

Supplier Data

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (AB97051)

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

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

The expression profile/ molecular weight observed is consistent with what has been described in the literature (PMID : 28541286 and PMID : 157438355).

Exposure time : Lane 1 : 180 seconds; Lanes 2-3 : 59 seconds; Lane 4 : 81 seconds; Lane 5 : 59 seconds

All lanes:

Western blot - Anti-PSAP antibody [RM1353] (ab324885) at 1/1000 dilution

Lane 1:

C6 (rat glial tumor glial cell) whole cell lysate at 20 µg

Lane 2:

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

Lane 3:

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

Lane 4:

SH-SY5Y (human neuroblastoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

PC-12 (rat adrenal gland pheochromocytoma cell) whole cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Observed band size: 60-80 kDa,12 kDa

false