Recombinant Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-321279-anti-erbb-2-antibody-epr19547-12-immunohistochemistry.jpg "Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)")
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR19547-12] to ErbB2 / HER2 - BSA and Azide free
- Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, IP, Indirect ELISA
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free
See all ErbB2 / HER2 primary antibodies -
Description
Rabbit monoclonal [EPR19547-12] to ErbB2 / HER2 - BSA and Azide free -
Host species
Rabbit -
Specificity
This antibody could recognize ErbB2 and weak cross reaction with ErbB4.
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Tested applications
Suitable for: Flow Cyt (Intra), IHC-P, ICC/IF, IP, Indirect ELISAmore details
Unsuitable for: WB -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- IHC-P: Human breast cancer tissue. ICC/IF: SK-BR-3 cells. Flow Cyt (intra): SK-BR-3 cells. IP: HeLa whole cell lysate.
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General notes
ab222482 is the carrier-free version of ab214275.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C. Do Not Freeze. -
Storage buffer
pH: 7.2
Constituent: PBS -
Carrier free
Yes -
Concentration information loading... -
Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR19547-12 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Conjugation kits
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab222482 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 |
|---|---|---|
| Flow Cyt (Intra) |
Use at an assay dependent concentration.
ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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| IHC-P |
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
|
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| ICC/IF |
Use at an assay dependent concentration.
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| IP |
Use at an assay dependent concentration.
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| Indirect ELISA |
Use at an assay dependent concentration.
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| Notes |
|---|
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Flow Cyt (Intra)
Use at an assay dependent concentration. ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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IHC-P
Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
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ICC/IF
Use at an assay dependent concentration. |
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IP
Use at an assay dependent concentration. |
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Indirect ELISA
Use at an assay dependent concentration. |
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: 13866 Mouse
- Entrez Gene: 24337 Rat
- Omim: 164870 Human
- SwissProt: P04626 Human
- SwissProt: P70424 Mouse
- SwissProt: P06494 Rat
- Unigene: 446352 Human
see all -
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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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)
Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ErbB2 / HER2 with ab214275 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane staining on Human breast cancer is observed [PMID: 18437174].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214275).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-321278-anti-erbb-2-antibody-epr19547-12-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)Immunohistochemical analysis of paraffin-embedded Human breast cancer tissue labeling ErbB 2 with ab214275 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Membrane staining on Human breast cancer is observed [PMID: 18437174].
Counter stained with Hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is ab97051 at 1/500 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214275).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-321277-anti-erbb-2-antibody-epr19547-12-immunohistochemistry.jpg)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)Immunohistochemical analysis of paraffin-embedded Human breast tissue labeling ErbB2 / HER2 with ab214275 at 1/4000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.
Negative control: No staining on normal Human breast [PMID: 15150568].
Counter stained with Hematoxylin.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214275).
Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
-
![Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-321276-anti-erbb-2-antibody-epr19547-12-immunofluorescence.jpg)
Immunocytochemistry/ Immunofluorescence - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)Immunofluorescent analysis of 100% methanol-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling ErbB2 / HER2 with ab214275 at 1/250 dilution, followed by Goat Anti-Rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).
Confocal image showing membrane staining on SK-BR-3 cells.
The nuclear counterstain is DAPI (blue).
Tubulin is detected with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (red).
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214275).
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![Flow Cytometry (Intracellular) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-321275-anti-erbb-2-antibody-epr19547-12-flow-cytometry.jpg)
Flow Cytometry (Intracellular) - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed SK-BR-3 (Human mammary gland adenocarcinoma cell line) cells labeling ErbB2 / HER2 with ab214275 at 1/60 dilution (red) compared with a RabbitIgG,monoclonal [EPR25A] - Isotype control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214275).
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![Immunoprecipitation - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-321274-anti-erbb-2-antibody-epr19547-12-immunoprecipitation.jpg)
Immunoprecipitation - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)ErbB2 / HER2 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab214275 at 1/30 dilution.
Western blot was performed from the immunoprecipitate using ab214275 at 1/1000 dilution.
VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution.
Lane 1: HeLa whole cell lysate 10µg (Input).
Lane 2: ab214275 IP in HeLa whole cell lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab214275 in HeLa whole cell lysate.
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 3 seconds.
This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab214275).
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![Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-487752-antierbb2her2antibodywesternblotrecominanthuman.jpg)
Western blot - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)All lanes : Anti-ErbB2 / HER2 antibody [EPR19547-12] (ab214275) at 1/1000 dilution
Lane 1 : His-tagged Recombinant Human ErbB2 protein (aa676-1255)
Lane 2 : His-tagged Recombinant Human ErbB4 protein (aa676-1308)
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 137 kDa
Observed band size: 80,90 kDa why is the actual band size different from the predicted?This data was developed using the same antibody clone in a different buffer formulation (ab222482).
Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 180 seconds.
This antibody could recognize ErbB2 and weak cross reaction with ErbB4.
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![Indirect ELISA - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-487753-antierbb2her2antibodyelisa.jpg)
Indirect ELISA - Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)This data was developed using the same antibody clone in a different buffer formulation (ab222482).
ELISA using ab214275 at using 1000 ng/ml an antigen concentration range of 500-0 ng/ml for Human ERBB2 and ERBB4 Recombinant Proteins. Goat Anti-Rabbit IgG (HRL) (1/50000) was used as the secondary antibody. 3,3’,5,5’-Tetramethylbenzidine (TMB) used as substrate solution.
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![Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)](https://www.abcam.com/ps/products/222/ab222482/Images/ab222482-7-benefits-of-recombinant-antibodies.png)
Anti-ErbB2 / HER2 antibody [EPR19547-12] - BSA and Azide free (ab222482)
Protocols
Datasheets and documents
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Datasheet download
Certificate of Compliance
References (1)
ab222482 has been referenced in 1 publication.
- Chen M et al. A Novel NIR Fluorescent Nanoprobe Targeting HER2-Positive Breast Cancer: Tra-TTR-A. Bioinorg Chem Appl 2021:2495958 (2021). PubMed: 35003236
