Anti-WDR4 antibody [EPR11052]

• IHC-P

Supplier Data

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR4 antibody [EPR11052] (AB169526)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human prostatic hyperplasia tissue sections labeling WDR4 with purified ab169526 at 1 : 1000 (1.026 μg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-WDR4 antibody [EPR11052] (AB169526)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma tissue sections labeling WDR4 with purified ab169526 at 1 : 1000 (1.026 μg/ml). Heat mediated antigen retrieval was performed using Bond™ Epitope Retrieval Solution 1 (pH 6.0). Tissue was counterstained with Hematoxylin. Rabbit specific IHC polymer detection kit HRP/DAB (ab209101) was used. PBS instead of the primary antibody was used as the negative control. The immunostaining was performed on a Leica Biosystems BOND^® RX instrument.

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-WDR4 antibody [EPR11052] (AB169526)

Flow Cytometry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labelling WDR4 with purified ab169526 at 1/100 dilution (10 µg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150081) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (black). Unlabelled control - Cell without incubation with primary antibody and secondary antibody (blue).

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-WDR4 antibody [EPR11052] (AB169526)

Immunocytochemistry analysis of HeLa (Human cervix adenocarcinoma epithelial cell) cells labeling WDR4 with purified ab169526 at 1 : 100 dilution (10.2 µg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-100. Cells were counterstained with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (2.5 µg/ml) (ab195889) (red). Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) was used as the secondary antibody at 1 : 1000 (2 µg/ml) dilution. DAPI (blue) was used as a nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• WB

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Western blot - Anti-WDR4 antibody [EPR11052] (AB169526)

All lanes:

Western blot - Anti-WDR4 antibody [EPR11052] (ab169526) at 1/1000 dilution

Lane 1:

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

Lane 2:

HeLa (Human cervix adenocarcinoma epithelial cell) whole cell lysate at 20 µg

Lane 3:

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

Lane 4:

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

Secondary

All lanes:

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

Predicted band size: 45 kDa

Observed band size: 45 kDa

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• WB

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Western blot - Anti-WDR4 antibody [EPR11052] (AB169526)

Lanes 1-3 : Merged signal (red and green). Green - ab169526 observed at 49 kDa. Red - loading control ab8245 observed at 36 kDa.

ab169526 Anti-WDR4 antibody [EPR11052] was shown to specifically react with WDR4 in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265109 (knockout cell lysate ab258285) was used. Wild-type and WDR4 knockout samples were subjected to SDS-PAGE. ab169526 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

All lanes:

Western blot - Anti-WDR4 antibody [EPR11052] (ab169526) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 20 µg

Lane 2:

WDR4 knockout HeLa cell lysate at 20 µg

Lane 2:

Western blot - Human WDR4 knockout HeLa cell line (<a href='/en-us/products/cell-lines/human-wdr4-knockout-hela-cell-line-ab265109'>ab265109</a>)

Lane 3:

HepG2 cell lysate at 20 µg

Secondary

All lanes:

Western blot - Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-irdye-800cw-preadsorbed-ab216773'>ab216773</a>) at 1/10000 dilution

Predicted band size: 45 kDa

Observed band size: 49 kDa

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