Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling ErbB2 / HER2 with ab16901 at 1/400 dilution. the tissue was fixed with formaldehyde; heat mediated antigen retrieval was performed using a acetate buffer pH6. An undiluted goat anti-mouse HRP conjugated seconday antibody was used.

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Anti-ErbB2 / HER2 antibody [3B5] (ab16901) at 1/250 dilution + Human Huh7 whole cell lysate at 20 µg

Secondary
Goat anti-mouse HRP conjugate at 1/5000 dilution

Predicted band size: 138 kDa
Observed band size: 138 kDa
Additional bands at: 48 kDa (possible non-specific binding)


Exposure time: 5 minutes

Blocking: 5% milk for 1 hour at 25°C.

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Mamma tumor Her2Neu tissue labeling ErbB 2 with ab16901.

Anti-ErbB2 / HER2 antibody [3B5] (ab16901) at 1 µg/ml + Whole cell lysate HEK293

Predicted band size: 138 kDa

Overlay histogram showing LoVo cells stained with ab16901 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16901, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x10⁶ cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in LoVo cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma labeling ErbB 2 with ab16901 at 1/100.

Immunohistochemistry analysis of formalin fixed, paraffin embedded tissue section of human breast carcinoma labeling ErbB 2 with at16901 at dilution of 1/1000.