The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/500 - 1/1000. Detects a band of approximately 185 kDa (predicted molecular weight: 138 kDa).
ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
Is unsuitable for ICC/IF,IHC-Fr or IHC-P.
Protein tyrosine kinase that is part of several cell surface receptor complexes, but that apparently needs a coreceptor for ligand binding. Essential component of a neuregulin-receptor complex, although neuregulins do not interact with it alone. GP30 is a potential ligand for this receptor. Regulates outgrowth and stabilization of peripheral microtubules (MTs). Upon ERBB2 activation, the MEMO1-RHOA-DIAPH1 signaling pathway elicits the phosphorylation and thus the inhibition of GSK3B at cell membrane. This prevents the phosphorylation of APC and CLASP2, allowing its association with the cell membrane. In turn, membrane-bound APC allows the localization of MACF1 to the cell membrane, which is required for microtubule capture and stabilization. In the nucleus is involved in transcriptional regulation. Associates with the 5'-TCAAATTC-3' sequence in the PTGS2/COX-2 promoter and activates its transcription. Implicated in transcriptional activation of CDKN1A; the function involves STAT3 and SRC. Involved in the transcription of rRNA genes by RNA Pol I and enhances protein synthesis and cell growth.
Expressed in a variety of tumor tissues including primary breast tumors and tumors from small bowel, esophagus, kidney and mouth.
Involvement in disease
Hereditary diffuse gastric cancer Glioma Ovarian cancer Lung cancer Gastric cancer Chromosomal aberrations involving ERBB2 may be a cause gastric cancer. Deletions within 17q12 region producing fusion transcripts with CDK12, leading to CDK12-ERBB2 fusion leading to truncated CDK12 protein not in-frame with ERBB2.
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily. Contains 1 protein kinase domain.
Autophosphorylated. Autophosphorylation occurs in trans, i.e. one subunit of the dimeric receptor phosphorylates tyrosine residues on the other subunit (Probable). Ligand-binding increases phosphorylation on tyrosine residues (PubMed:27134172). Signaling via SEMA4C promotes phosphorylation at Tyr-1248 (PubMed:17554007). Dephosphorylated by PTPN12 (PubMed:27134172).
Cytoplasm. Nucleus and Cell membrane. Cytoplasm, perinuclear region. Nucleus. Translocation to the nucleus requires endocytosis, probably endosomal sorting and is mediated by importin beta-1/KPNB1.
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
For western blottingab53290 at 1/200 dilution and ab131366 VeriBlot for IP (HRP) at 1/1000 was used as the secondary antibody.
Blocking and Diluting buffer and concentration: 5% NFDM/TBST.
Dot Blot - Anti-ErbB 2 (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.
Western blot - Anti-ErbB 2 (phospho Y1139) antibody [EP1046Y] (ab53290)
All lanes : Anti-ErbB 2 (phospho Y1139) antibody [EP1046Y] (ab53290) at 1/1000 dilution
Lane 1 : SKBR3 cell lysate Lane 2 : SKBR3 cell lysate - treated with EGF
Lysates/proteins at 10 µg per lane.
Secondary All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution
Western blot - Anti-ErbB 2 (phospho Y1139) antibody [EP1046Y] (ab53290)This image is courtesy of an Abreview by Surekha Pimple.
All lanes : Anti-ErbB 2 (phospho Y1139) antibody [EP1046Y] (ab53290) at 1/2000 dilution
Lane 1 : HTB-2O Human breast ductal carcinoma lysate untreated Lanes 2-9 : HTB-2O Human breast ductal carcinoma lysate treated with HER-2 inhibitor
Lysates/proteins at 10 µg per lane.
Secondary All lanes : HRP-conjugated Mouse anti-Rabbit monoclonal at 1/20000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 138 kDa
Exposure time: 5 seconds
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.