Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] - BSA and Azide free

• ICC/IF

Supplier Data

Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] - BSA and Azide free (AB245120)

Immunofluorescent analysis of 100% methanol fixed SK-BR-3 (human breast adenocarcinoma epithelial cell) cells labeling ErbB2 / HER2 with ab241325 at 1/50 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

Confocal image showing membranous staining in SK-BR-3 cells.

Negative control : MCF7 (Human breast adenocarcinoma cell line) (PMID : 20697531). Negative staining was obtained from 4% PFA fixation.

The nuclear counter stain is DAPI (blue). Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor^® 594) at a 1/200 dilution (red).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241325).

• WB

Lab

Western blot - Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] - BSA and Azide free (AB245120)

False colour image of Western blot : Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab241325 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type HeLa cell lysates with no signal observed at this size in ERBB2 knockout cell line ab255387 (knockout cell lysate ab263758). To generate this image, wild-type and ERBB2 knockout HeLa 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 (IRDye^® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed (ab216776) at 1/20000 dilution.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241325).

All lanes:

Western blot - Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] (<a href='/en-us/products/primary-antibodies/erbb2-her2-antibody-cpt-r322-27-12-ab241325'>ab241325</a>) at 1/1000 dilution

Lane 1:

Wild-type HeLa cell lysate at 40 µg

Lane 2:

ERBB2 knockout HeLa cell lysate at 40 µg

Predicted band size: 137 kDa

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• ELISA

Supplier Data

ELISA - Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] - BSA and Azide free (AB245120)

ELISA - Anti-ErbB2 / HER2 antibody [CPT-R32.2-27-12] (ab241325) used at 1000 ng/mL.

50 ng CPTAC-36d (1000 ng/mL, 50 μL per well) was coated onto 96-wells. Serial dilutions (0, 0.2, 0.9, 3.9, 15, 62, 250, 1000 ng/mL) of ab241325 (50 μL) were incubated with CPTAC-36d in each well for 60 minutes by shaking. This was followed by adding goat anti-rabbit IgG, (H+L), phosphatase-conjugated secondary antibody (50 μL, 1/2500) into each well and incubating for another 40 minutes. After washing, 50 μL of PNPP was added and incubated for 15 minutes without shaking for color development. OD was read at 405nm within 5 minutes.

Primary antibody concentration range : 0.01 - 1.0 μg/mL.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab241325).