Anti-EGFR (phospho Y992) antibody

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y992) antibody (AB5638)

Immunohistochemistry analysis of EGFR (pY992) with ab5638 at 1/20 dilution showing staining in the cytoplasm and membrane of paraffin-embedded human lung adenocarcinoma (right) compared to a negative control without primary antibody (left).

Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min.

• IHC-P

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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-EGFR (phospho Y992) antibody (AB5638)

Immunohistochemistry analysis of EGFR (pY992) with ab5638 at 1/20 dilution showing staining in the cytoplasm and membrane of paraffin-embedded human hepatocellular carcinoma (right) compared to a negative control without primary antibody (left).

Antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min.

• ICC

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Immunocytochemistry - Anti-EGFR (phospho Y992) antibody (AB5638)

Immunocytochemical analysis of EGFR [pY992] was performed using A-431 cells treated with 200 ng/mL of EGF for 10 minutes. The cells were fixed with 4% paraformaldehyde, permeabilized with 0.1% Triton™ X-100, and blocked with 1% BSA. The cells were labeled with ab5638 at 1/250 dilution and incubated overnight at 4°C followed by a Goat anti-Mouse IgG (H+L) Secondary Antibody, Alexa Fluor® 488 conjugate at a dilution of 1/2000 for 45 minutes at room temperature (Panel a : green). Nuclei (Panel b : blue) DAPI (Panel c : red) F-actin was stained with Rhodamine Phalloidin at 1/300. Panel d represents the merged image showing membranous localization. Panel e represents cells treated with antagonist, Afatinib (1μM for 6hrs) followed by EGF (200 ng/mL for 10 minutes), showing reduced Phospho-EGFR staining. Panel f shows untreated cells with no signal. Panel g represents control cells with no primary antibody to assess background.

• WB

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Western blot - Anti-EGFR (phospho Y992) antibody (AB5638)

All lanes:

Western blot - Anti-EGFR (phospho Y992) antibody (ab5638) at 1/1000 dilution

Lane 1:

A431 cells unstimulated

Lanes 2 - 5:

A431 cells stimulated with 200 ng/mL EGF for 15 minutes

Predicted band size: 134 kDa

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