Immunocytochemistry/ Immunofluorescence analysis of A431 (human epidermoid carcinoma epithelial cell) cells labeling EGFR with purified ab223499 at 1:100 (4.3 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

Flow Cytometry analysis of A431 cells treated with 100ng/ml EGF for 10mins cells labeling EGFR with purified ab223499 at 1:4200 dilution (0.10 µg/ml) - Red. Cells were fixed with 4% paraformaldehyde . A Goat anti rabbit IgG (Alexa Fluor® 488, ab150081) secondary antibody was used at 1:2000 dilution. Isotype control - Rabbit monoclonal IgG (ab172730) - Black. Unlabeled control - Blue. Untreated cells - Green

All lanes : Anti-EGFR (phospho Y1068) antibody [SP353] (ab223499) at 1/400 dilution

Lane 1 : A431 (human epidermoid carcinoma cell line) + EGF cell lysate
Lane 2 : A431 (human epidermoid carcinoma cell line) cell lysate

PFA-fixed, paraffin-embedded human lung cancer tissue stained for EGFR using ab223499 at 1/100 dilution in immunohistochemical analysis.

 

PFA-fixed, paraffin-embedded human lung squamous cell carcinoma tissue stained for EGFR using ab223499 at 1/100 dilution in immunohistochemical analysis.

Flow cytometric analysis of ab223499 in A431 (human epidermoid carcinoma cell line) + EGF cells (green) at 1/400 dilution compared to negative control of rabbit IgG (blue).