Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y]

• IP

Lab

Immunoprecipitation - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (AB53290)

ab53290 at 1/70 dilution immunoprecipitating ErbB 2 (phospho Y1139) in A431(human epidermoid carcinoma) whole cell lysate.

Lane 1 (input) : A431treated with 100 ng/mL EGF for 10 minutes whole cell lysate 10μg

Lane 2 (+) : ab53290 + A431 treated with 100 ng/mL EGF for 10 minutes whole cell lysate

Lane 3 (-) : Rabbit monoclonal IgG (ab172730) instead of ab53290 in A431treated with 100 ng/mL EGF for 10 minutes whole cell lysate

VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.

Blocking and Diluting buffer and concentration : 5% NFDM/TBST.

All lanes:

Immunoprecipitation - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (ab53290)

Predicted band size: 137 kDa

Observed band size: 185 kDa

false

Exposure time: 5s

• WB

Lab

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (AB53290)

Blocking and dilution buffer : 5% NFDM/TBST.

All lanes:

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (ab53290) at 1/500 dilution

Lane 1:

A431 whole cell lysate - untreated at 10 µg

Lane 2:

A431 whole cell lysate - treated with Epidermal Growth Factor (EGF) at 10 µg

Secondary

All lanes:

Peroxidase-conjugated goat anti-rabbit IgG (H+L) at 1/1000 dilution

Predicted band size: 137 kDa

Observed band size: 185 kDa

false

Exposure time: 3min

• WB

Unknown

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (AB53290)

All lanes:

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (ab53290) at 1/1000 dilution

Lane 1:

SKBR3 cell lysate at 10 µg

Lane 2:

SKBR3 cell lysate - treated with EGF at 10 µg

Secondary

All lanes:

HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

Predicted band size: 137 kDa

Observed band size: 185 kDa

false

• WB

AbReview21787****

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (AB53290)

Lanes 2-9 are treated with an increasing concentration of HER2 inhibitor.
Blocking performed with 5% Milk for 1 hour at room temperature.
Antibody diluted in CPPT and incubated for 16 hours at 4°C.
Performed under denaturing conditions.

All lanes:

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (ab53290) at 1/2000 dilution

Lane 1:

HTB-2O Human breast ductal carcinoma lysate untreated at 10 µg

Lanes 2 - 9:

HTB-2O Human breast ductal carcinoma lysate treated with HER-2 inhibitor at 10 µg

Secondary

All lanes:

HRP-conjugated Mouse anti-Rabbit monoclonal at 1/20000 dilution

Predicted band size: 137 kDa

true

Exposure time: 5s

This image is courtesy of an Abreview by Surekha Pimple.

• Dot

Unknown

Dot Blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (AB53290)

Dot blot analysis of ErbB 2 (pY1139) peptide (Lane 1) and ErbB 2 non-phospho peptide (Lane 2) labelling ErbB 2 (phospho Y1139) with ab53290 at a dilution of 1/1000. A Peroxidase-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

Blocking and dilution buffer : 5% NFDM/TBST.

Exposure time : 3 minutes.

• WB

CiteAb

Western blot - Anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] (AB53290)

ErbB2 / HER2 (phospho Y1139) western blot using anti-ErbB2 / HER2 (phospho Y1139) antibody [EP1046Y] ab53290. Publication image and figure legend from Li, J., Ma, M., et al., 2020, Mol Cancer, PubMed 32917240.

ab53290 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 ab53290 please see the product overview.

HER2–103 promoted EGFR/HER3 interaction and activation in TNBC. a Circ-HER2 shRNAs or scramble shRNA stably transfected MDA-MB-231 or MDA-MB-468 cells, as well as circ-HER2 and circ-HER2 noATG vector stably transfected MDA-MB-231 or MDA-MB-468 cells were subjected to immunoblot with indicated antibodies. The transfection efficiency of above cells was verified by q-PCR. b circ-HER2 or circ-HER2 noATG vector stably transfected BT549 cells were subjected to immunoblot with indicated antibodies. The transfection efficiency was verified both by q-PCR. c Immunoprecipitation using HER3 or EGFR antibodies were performed in circ-HER2 shRNAs or scramble shRNA stably transfected MDA-MB-468 cells with indicated antibodies. d Left, circ-HER2 was stably transfected to MDA-MB-468 cells, HER2–103 was immunoprecipitated, followed by immunoblot with antibodies against EGFR and HER3; middle, EGFR and HER3 were immunoprecipitated in above mentioned cells, followed by immunoblot with antibodies against HER2–103. Right, circ-HER2 was stably transfected to MDA-MB-231 cells, HER2–103 or EGFR were immunoprecipitated followed by immunoblot with antibodies against EGFR and HER2–103. e Upper left, EGFR domains, namely, L1, CR1, L2, CR2, juxta-membrane (JM) segment and internal cellular domain (ICD). Upper right, HA-tagged EGFR domains and Flag-tagged HER2–103 were co-transfected into 293 T cells. Coimmunoprecipitated truncated EGFR protein was detected by anti-HA antibody after immunoprecipitation with anti-Flag antibody. Lower left, HER3 domains, namely, L1, CR1, L2, CR2, juxta-membrane (JM) segment and internal cellular domain (ICD). HA-tagged HER3 domains and Flag tagged HER2–103 were co-transfected into HEK293T cells. Coimmunoprecipitated truncated HER3 protein was detected by anti-HA antibody after immunoprecipitation with anti-Flag antibody. f HER2–103 was dose-dependently transfected into MDA-MB-468 cells. Interaction between EGFR/HER3 was determined mutually by immunoprecipitation, with indicated antibodies. g The kinase activity of EGFR pY1068 was detected following circ-HER2 stably transfection in MDA-MB-468 cells. Lines show the mean ± SD, ***, p < 0.001; **, p < 0.01; *p < 0.05. Data are representative from at least 2–3 experiments with similar results

false