Anti-EGFR (phospho S1142) antibody

• ICC/IF

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Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho S1142) antibody (AB76194)

ab76194, at a 1/200 dilution, staining EGFR in A431 cells by Immunoflurescence.
Left image : untreated.
Middle image : treated with Calyculin A (50 nM) for 15 minutes.
Right image : treated with Calyculin A (50 nM) for 15 minutes with blocking peptide, phospho EGFR (Ser-1142).

• WB

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Western blot - Anti-EGFR (phospho S1142) antibody (AB76194)

All lanes:

Western blot - Anti-EGFR (phospho S1142) antibody (ab76194) at 1/1000 dilution

Lane 1:

A431 cells treated with Calyculin A (100 nM) for 30 minutes

Lane 2:

A431 cells treated with Calyculin A (100 nM) for 30 minutes and then treated with lambda phosphatase

Predicted band size: 134 kDa

Observed band size: 180 kDa

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• WB

CiteAb

Western blot - Anti-EGFR (phospho S1142) antibody (AB76194)

EGFR (phospho S1142) western blot using anti-EGFR (phospho S1142) antibody ab76194. Publication image and figure legend from Jones, S., King, P. J., et al., 2020, Sci Rep, PubMed 31959764.

ab76194 was used in this publication in western blot. This may not be the same as the application(s) guaranteed by Abcam. For a full list of applications guaranteed by Abcam for ab76194 please see the product overview.

Sym004-mediated endocytosis does not require EGFR TK activity and only partially depends on clathrin. (a) Serum-starved SCC47 cells were treated with cetuximab, Sym004 or EGF, then lysed. (b) SCC47 cells were serum-starved, final 30 min in the presence of erlotinib (0.5 μM). The cells were treated with Sym004 or EGF for 6 h, then lysed. EGFR degradation was quantified as remaining EGFR (normalised to tubulin) relative to untreated cells. (c) The cells treated as in (b) were fixed and stained with EGFR-AF488 and Phalloidin-AF647 antibodies. (d) Serum-starved, clathrin heavy chain-depleted (or control) SCC47 cells were treated with Sym004 (3 μg/ml or 30 μg/ml) or EGF for 6 h, then lysed. EGFR degradation was quantified as remaining EGFR (normalised to tubulin) relative to untreated cells. (e) The cells treated as in (d) were fixed and stained with EGFR-AF488. (f) SCC47 cells were serum-starved, final 30 min in the presence of cytochalasin D (5 μM or 50 μM) or DMSO (control). The cells were treated with Sym004 or EGF for 6 h, then lysed. EGFR degradation was quantified as remaining EGFR (normalised to tubulin) relative to untreated cells. Error bars, SEM, *p < 0.05. Nuclei were stained with Hoechst 33342. All immunoblots were cropped for clarity.

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