Anti-EGFR (phospho Y1173) antibody [E124]

• ICC/IF

Lab

Immunocytochemistry/ Immunofluorescence - Anti-EGFR (phospho Y1173) antibody [E124] (AB32578)

Immunocytochemistry analysis of A431 (Human epidermoid carcinoma epithelial cell) treated with EGF(100ng/ml 5min) cells labeling EGFR with Purified ab32578 at 1 : 2000 dilution (0.1 μg/ml). Cells were fixed in 4% Paraformaldehyde and permeabilized with 0.1% tritonX-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 (blue) was used as nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

• Flow Cyt (Intra)

Unknown

Flow Cytometry (Intracellular) - Anti-EGFR (phospho Y1173) antibody [E124] (AB32578)

Intracellular Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) treated (Red)/untreated (Green) with 500ng/ml Trichostatin A for 4 hours with purified ab177178 at 1/1300 dilution. The secondary antibody was Goat anti rabbit IgG (Alexa Fluorr^® 488) at 1/2000 dilution. A Rabbit monoclonal IgG (Black) was used as the isotype control and cells without incubation with primary antibody and secondary antibody (Blue) were used as unlabeled control.

• WB

Supplier Data

Western blot - Anti-EGFR (phospho Y1173) antibody [E124] (AB32578)

All lanes:

Western blot - Anti-EGFR (phospho Y1173) antibody [E124] (ab32578) at 1/1000 dilution

Lane 1:

Untreated A431 (Human epidermoid carcinoma epithelial cell) whole cell lysate at 15 µg

Lane 2:

A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30 minutes whole cell lysate at 15 µg

Lane 3:

A431 (Human epidermoid carcinoma epithelial cell) treated with 100ng/ml EGF for 30 minutes whole cell lysate, then the membrane treated with Alkaline Phosphatase for 1 hour at 15 µg

Secondary

All lanes:

Western blot - Goat Anti-Rabbit IgG H&L (HRP) (<a href='/en-us/products/secondary-antibodies/goat-rabbit-igg-h-l-hrp-ab97051'>ab97051</a>) at 1/20000 dilution

Predicted band size: 134 kDa

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• WB

CiteAb

Western blot - Anti-EGFR (phospho Y1173) antibody [E124] (AB32578)

EGFR (phospho Y1173) western blot using anti-EGFR (phospho Y1173) antibody [E124] ab32578. Publication image and figure legend from Bönisch, M., Sellmann, C., et al., 2017, Protein Eng Des Sel, PubMed 28981885.

ab32578 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 ab32578 please see the product overview.

Biophysical and functional characterisation of B10v5 × hu225M with or without mutations in the CH1 : CL interface. Analytical SEC after protein A purification of bsAbs produced in (A) HEK293-6E or (B) bsAbs produced in ExpiCHO after preparative SEC. The area under the curve in percent of all detected peak areas is given in the graph. (C) Simultaneous binding of both antigens cMET and EGFR using biolayer interferometry. BsAbs (wt, MaB40, MaB5/40, MaB21/40, MaB45/40) or antibodies consisting of only one Fab (oa for ‘one-armed’) were allowed to bind to cMET coated biosensors. Subsequently, biosensors were incubated with EGFR. (D) Differential scanning calorimetry of wildtype, interface mutants and ‘one-armed’ constructs (Table III). (E) Western blot analysis of the inhibition of EGFR and cMET phosphorylation in A549 cells by bsAbs with our without interface designs.

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