Anti-EGFR (phospho Y845) antibody [EPR2149Y] - BSA and Azide free

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y845) antibody [EPR2149Y] - BSA and Azide free (AB240934)

Immunofluorescent analysis of 4% paraformaldehyde-fixed,0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cell lines labeling EGFR with ab97613 (purified) at 1/50 dilution,followed by Goat anti-Rabbit IgG H & L (Alexa Fluor^® 488) (ab150077) secondary antibody at 1/1000 dilution

Cells were counterstained with ab195889 (Anti-alpha Tubulin antibody [DM1A] ) - Microtubule Marker (Alexa Fluor^® 594)

The nuclear counterstain is DAPI (blue).

The green staining on the membrane was increased in the EGF (100 ng/ml, 10 min) treated A431 cells when compared with A431 cells without treatment. After LP treatment, the green signaling was obviously decreased.

For the pan antibody, there was no great difference after EGF (100 ng/ml, 10 min) or EGF (100 ng/ml, 10 min) + LP treatment. The data showed mostly membranous staining.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab97613)

• Flow Cyt (Intra)

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Flow Cytometry (Intracellular) - Anti-EGFR (phospho Y845) antibody [EPR2149Y] - BSA and Azide free (AB240934)

Intracellular Flow Cytometry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 10 minutes cells labeling EGFR with purified ab97613 at 1/400 dilution (1μg/ml) (red). Cells were fixed with 4% Paraformaldehyde and permeabilised with 90% Methanol. A Goat anti rabbit IgG (Alexa Fluor^® 488, ab150077) secondary antibody was used at 1/2000. Isotype control - Rabbit monoclonal IgG (Black). Unlabeled control - Cell without incubation with primary antibody and secondary antibody (Blue). Untreated A431 cells (Green).

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab97613).

• Dot

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Dot Blot - Anti-EGFR (phospho Y845) antibody [EPR2149Y] - BSA and Azide free (AB240934)

Dot Blot analysis.

Lane 1 : EGFR (pY845) phospho peptide.

Lane 2 : EGFR non-phospho peptide labeling EGFR (phospho Y845) with ab97613 (unpurified) at 1/1000 dilution.

5% NFDM/TBST was used as the diluting and blocking buffer. ab97051 Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as the secondary antibody at 1/100000 dilution. Exposure time : 3 minutes.

This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab97613)