Anti-CENPB antibody

• WB

Lab

Western blot - Anti-CENPB antibody (AB25734)

Lanes 1 - 4 : Merged signal (red and green). Green - ab25734 observed at 77 kDa. Red - loading control ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37 kDa.

ab25734 was shown to react with CENPB in wild-type A431 cells in Western blot with loss of signal observed in CENPB knockout sample.Wild-type A431 and CENPB knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3 % milk in TBS-T (0.1 % Tween^®) before incubation with ab25734 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at 1 μg/ml and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 h at room temperature before imaging.

All lanes:

Western blot - Anti-CENPB antibody (ab25734) at 1 µg/mL

Lane 1:

Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

CENPB knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate at 20 µg

Lane 2:

Western blot - Human CENPB knockout A-431 cell line (<a href='/en-us/products/cell-lines/human-cenpb-knockout-a-431-cell-line-ab274919'>ab274919</a>)

Lane 3:

Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate at 20 µg

Lane 4:

Daudi (Human Burkitt's lymphoma cell line) whole cell lysate at 20 µg

Predicted band size: 65 kDa

Observed band size: 77 kDa

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• ICC/IF

AbReview50051****

Immunocytochemistry/ Immunofluorescence - Anti-CENPB antibody (AB25734)

4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized HeLa (human epithelial cell line from cervix adenocarcinoma) cells stained for CENPB (red) using ab25734 at 1/200 dilution in ICC/IF.

The nuclear counterstain is DAPI (blue). Alpha-Tubulin is stained with an mouse monoclonal anti-alpha Tubulin antibody (green).

Image courtesy of an AbReview by Weiguo Zhang

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-CENPB antibody (AB25734)

ab25734 staining CENPB in HeLa cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab25734 at 1 μg/ml and ab7291, Mouse monoclonal [DM1A] to alpha Tubulin - Loading Control. Cells were then incubated with ab150120, Goat polyclonal Secondary Antibody to Mouse IgG - H&L (Alexa Fluor^® 594), pre-adsorbed at 1/1000 dilution (shown in green) and ab150081, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor^® 488), pre-adsorbed at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

Also suitable in cells fixed with 100% methanol (5 min).

Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

• ICC

AbReview9910****

Immunocytochemistry - Anti-CENPB antibody (AB25734)

ab25734 (1/1000) staining CENPB in HeLa cells (green). Cells were fixed with methanol and counterstained with DAPI in order to highlight the nucleus (red). Please refer to abreview for further experimental details.

This image is courtesy of an Abreview submitted by Dr Kirk McManus

• WB

Unknown

Western blot - Anti-CENPB antibody (AB25734)

All lanes:

Western blot - Anti-CENPB antibody (ab25734) at 1 µg/mL

Lane 1:

HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate at 20 µg

Lane 2:

Western blot - Jurkat whole cell lysate (<a href='/en-us/products/cell-lysates/jurkat-whole-cell-lysate-ab7899'>ab7899</a>) at 20 µg

Lane 3:

Western blot - A-431 whole cell lysate (<a href='/en-us/products/cell-lysates/a-431-whole-cell-lysate-ab7909'>ab7909</a>) at 20 µg

Secondary

All lanes:

Goat polyclonal to Rabbit IgG (Alexa Fluor® 680) at 1/10000 dilution

Predicted band size: 65 kDa

Observed band size: 65 kDa,74 kDa,75 kDa

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