重组Anti-KIAA0319L抗体[EPR28329-62]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

Immunohistochemical analysis of paraffin-embedded Human cerebrum tissue labeling KIAA0319L with ab315027 at 1/500 (1.038 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on human cerebrum. The section was incubated with ab315027 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

Immunohistochemical analysis of paraffin-embedded Mouse cerebrum tissue labeling KIAA0319L with ab315027 at 1/2000 (0.26 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on mouse cerebrum. The section was incubated with ab315027 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

Immunohistochemical analysis of paraffin-embedded Rat cerebrum tissue labeling KIAA0319L with ab315027 at 1/2000 (0.26 ug/ml) followed by a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection). Positive staining on rat cerebrum. The section was incubated with ab315027 for 30 mins at room temperature. The immunostaining was performed on a Leica Biosystems BOND® RX instrument. Counterstained with Hematoxylin.

Secondary antibody only control : Secondary antibody is a ready to use LeicaDS9800 (Bond™ Polymer Refine Detection).

Heat mediated antigen retrieval was performed with Tris-EDTA buffer (pH 9.0, Epitope Retrieval Solution2) for 20 mins.

• WB

Lab

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

Western blot : Anti-KIAA0319L antibody [EPR28329-62] ab315027 staining at 1/1000 dilution, shown in green; Mouse anti GAPDH ab8245 loading control staining at 1/20000 dilution, shown in magenta. A band was observed at 150 kDa in Wild-type HCT 116 cell lysates with no signal observed at this size in KIAA0319L knockout HCT 116 cell line. To generate this image, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 5% 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 800CW and Goat anti-Mouse 680RD at 1/20,000 dilution.

All lanes:

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (ab315027) at 1/1000 dilution

Lane 1:

Wild-type HCT 116 cell lysates at 20 µg

Lane 2:

KIAA0319L knockout HCT 116 cell lysates at 20 µg

Lane 3:

HeLa cell lysates at 20 µg

Lane 4:

HL-60 cell lysates at 20 µg

Secondary

All lanes:

Goat anti-Rabbit 800CW & Goat anti-Mouse 680RD at 1/20000 dilution

Predicted band size: 150 kDa

Observed band size: 150 kDa

false

• WB

Supplier Data

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

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

Negative control : HL-60 (PMID : 28679762).

The observed Mw of ~150KDa is higher than the predicted Mw due to glycosylation (PMID : 28679762).

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

All lanes:

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (ab315027) at 1/1000 dilution

Lane 1:

HeLa (human cervical adenocarcinoma epithelial cell) transfected with scrambled siRNA control whole cell lysate at 20 µg

Lane 2:

HeLa transfected with siRNA specifically targeting KIAA0319L whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

Huh7 (human hepatocellular carcinoma epithelial cell) whole cell lysate at 20 µg

Lane 5:

HL-60 (human acute promyelocytic leukemia promyeloblast) whole cell lysate at 20 µg

Lane 6:

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

Lane 7:

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

Lane 8:

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

Lane 9:

L6 (rat skeletal muscle myoblast) 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: 150 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

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

Deglycosylation induced a shift in the observed molecular weight from ~150KDa to ~100KDa (PMID : 28679762).

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

All lanes:

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (ab315027) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa whole cell lysate treated with Protein Deglycosylation MIX II 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: 150 kDa,100 kDa,36 kDa

false

Exposure time: 48s

• WB

Supplier Data

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (AB315027)

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

The observed Mw of ~150KDa is higher than the predicted Mw due to glycosylation (PMID : 28679762).

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

All lanes:

Western blot - Anti-KIAA0319L antibody [EPR28329-62] (ab315027) at 1/1000 dilution

Lane 1:

Human hippocampus tissue lysate at 20 µg

Lane 2:

Mouse brain tissue lysate at 20 µg

Lane 3:

Rat brain 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 1/100000 dilution

Observed band size: 150 kDa,36 kDa

false

Exposure time: 180s