Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)
Key features and details
- Rabbit polyclonal to ErbB2 / HER2 (phospho Y1248)
- Suitable for: WB, ELISA, IHC-P
- Reacts with: Human
- Isotype: IgG
Overview
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Product name
Anti-ErbB2 / HER2 (phospho Y1248) antibody
See all ErbB2 / HER2 primary antibodies -
Description
Rabbit polyclonal to ErbB2 / HER2 (phospho Y1248) -
Host species
Rabbit -
Tested applications
Suitable for: WB, ELISA, IHC-Pmore details -
Species reactivity
Reacts with: Human
Predicted to work with: Rat
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Immunogen
Synthetic peptide corresponding to Human ErbB2/ HER2 aa 1200 to the C-terminus (phospho Y1248) conjugated to keyhole limpet haemocyanin.
(Peptide available asab139514) -
Positive control
- This antibody gave a positive signal in EGF-treated A431 whole cell lysate. IHC-P: FFPE human breast adenocarcinoma tissue sections.
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General notes
Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.
Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.
We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.
In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.
We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.
Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.
Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.40
Preservative: 0.02% Sodium azide
Constituent: PBS
Batches of this product that have a concentration
Concentration information loading...Purity
Immunogen affinity purifiedClonality
PolyclonalIsotype
IgGResearch areas
Associated products
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Compatible Secondaries
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Isotype control
Applications
Our Abpromise guarantee covers the use of ab101229 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application Abreviews Notes WB Use a concentration of 1 µg/ml. Detects a band of approximately 180 kDa (predicted molecular weight: 138 kDa). ELISA Use at an assay dependent concentration. IHC-P Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. Target
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Function
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. -
Tissue specificity
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. -
Sequence similarities
Belongs to the protein kinase superfamily. Tyr protein kinase family. EGF receptor subfamily.
Contains 1 protein kinase domain. -
Post-translational
modificationsAutophosphorylated. 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). -
Cellular localization
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. - Information by UniProt
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Database links
- Entrez Gene: 2064 Human
- Entrez Gene: 24337 Rat
- Omim: 164870 Human
- SwissProt: P04626 Human
- SwissProt: P06494 Rat
- Unigene: 446352 Human
- Unigene: 93966 Rat
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Alternative names
- Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog antibody
- C erb B2/neu protein antibody
- CD340 antibody
see all
Images
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Western blot - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)All lanes : Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229) at 1 µg/ml
Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : EGF-Stimulated A431 Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 4 : EGF-Stimulated A431 Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 5 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lane 6 : EGF-Stimulated A431 Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 138 kDa
Exposure time: 3 minutesErbB2 / HER2 (phospho Y1248)contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)IHC image of ab101229 staining in FFPE human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab101229, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
ELISA - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)ab101229 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.
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Western blot - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)This image is courtesy of an anonymous AbreviewAll lanes : Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229) at 1/500 dilution
Lane 1 : Schwann cell extract untreated
Lane 2 : Schwann cell extract treated with 10 ng/ml GGF for 30 minutes.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 138 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteLanes on the left hand side show phosphorylation of ErbB2 / HER2 at tyrosine 1248 (ab101299) in rat Schwann cells upon incubation with 10 ng/ml GGF for 30 minutes (lane 2).
Treatment with GGF also induces phosphorylation of ErbB2 at tyrosine 877, which is observed on the right hand side blots.
Protocols
Datasheets and documents
References (1)
ab101229 has been referenced in 1 publication.
- Secombe KR et al. Targeting neratinib-induced diarrhea with budesonide and colesevelam in a rat model. Cancer Chemother Pharmacol N/A:N/A (2018). PubMed: 30535958
Images
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Western blot - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)All lanes : Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229) at 1 µg/ml
Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : EGF-Stimulated A431 Whole Cell Lysate
Lane 3 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 4 : EGF-Stimulated A431 Whole Cell Lysate with Immunizing peptide at 1 µg/ml
Lane 5 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lane 6 : EGF-Stimulated A431 Whole Cell Lysate with Non modified peptide at 1 µg/ml
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 138 kDa
Exposure time: 3 minutesErbB2 / HER2 (phospho Y1248)contains a number of potential glycosylation sites (SwissProt) which may explain its migration at a higher molecular weight than predicted.
-
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)IHC image of ab101229 staining in FFPE human breast adenocarcinoma formalin fixed paraffin embedded tissue section, performed on a Leica BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab101229, 1µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times. -
ELISA - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229)
ab101229 was tested using an Indirect ELISA approach. The wells were coated with peptide (1µg/ml at 100µl/well) overnight at 4°C, followed by a 5% BSA blocking step for 1 hour at room temperature. The primary Ab was then added at a dilution range of 1- 0.00025µg/ml (100µl/well) for 1hr at room temperature. A HRP-conjugated anti-rabbit IgG (heavy and light chain) was used as a secondary antibody at 1:20,000 dilution for 1hr at room temperature.
-
Western blot - Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229) This image is courtesy of an anonymous AbreviewAll lanes : Anti-ErbB2 / HER2 (phospho Y1248) antibody (ab101229) at 1/500 dilution
Lane 1 : Schwann cell extract untreated
Lane 2 : Schwann cell extract treated with 10 ng/ml GGF for 30 minutes.
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP conjugated goat anti-rabbit IgG at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Predicted band size: 138 kDa
Observed band size: 200 kDa why is the actual band size different from the predicted?
Exposure time: 1 minuteLanes on the left hand side show phosphorylation of ErbB2 / HER2 at tyrosine 1248 (ab101299) in rat Schwann cells upon incubation with 10 ng/ml GGF for 30 minutes (lane 2).
Treatment with GGF also induces phosphorylation of ErbB2 at tyrosine 877, which is observed on the right hand side blots.
