Recombinant Anti-ErbB2 / HER2 antibody [CAL27] - BSA and Azide free (ab251602)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [CAL27] to ErbB2 / HER2 - BSA and Azide free
- Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra)
- Knockout validated
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-ErbB2 / HER2 antibody [CAL27] - BSA and Azide free
See all ErbB2 / HER2 primary antibodies -
Description
Rabbit monoclonal [CAL27] to ErbB2 / HER2 - BSA and Azide free -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WB, ICC/IF, IP, Flow Cyt (Intra)more details -
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
- WB: HeLa, 4T1, C6, Wild-type HCT 116, Wild-type A549 and SK-BR-3 whole cell lysates. IHC-P: Human breast carcinoma 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
ab251602 is the carrier-free version of ab237715.
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.
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...
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Purity
Protein A purified -
Purification notes
Purity is greater than 99%. -
Clonality
Monoclonal -
Clone number
CAL27 -
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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Immunohistochemistry kits
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Isotype control
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KO cell lines
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab251602 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 |
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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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WB |
Use at an assay dependent concentration. Detects a band of approximately 180 kDa (predicted molecular weight: 137 kDa).
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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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Flow Cyt (Intra) |
Use at an assay dependent concentration.
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Notes |
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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. |
WB
Use at an assay dependent concentration. Detects a band of approximately 180 kDa (predicted molecular weight: 137 kDa). |
ICC/IF
Use at an assay dependent concentration. |
IP
Use at an assay dependent concentration. |
Flow Cyt (Intra)
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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All lanes : Anti-ErbB2 / HER2 antibody [CAL27] (ab237715) at 1/500 dilution
Lane 1 : Wild-type MCF7 cell lysate at 32 µg
Lane 2 : ERBB2 knockout MCF7 cell lysate at 32 µg
Lane 3 : SK-BR-3 cell lysate at 16 µg
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 180 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 containing PBS and sodium azide (ab237715).
Anti-ErbB2 / HER2 antibody [CAL27] staining at 1/500 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab237715 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type MCF7 cell lysates with no signal observed at this size in ERBB2 knockout cell line ab286260 (knockout cell lysate AB300208).
To generate this image, wild-type and ERBB2 knockout MCF7 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were HRP conjugated Goat anti-Rabbit (H+L) and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
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All lanes : Anti-ErbB2 / HER2 antibody [CAL27] (ab237715) at 1/1000 dilution
Lane 1 : Wild-type A549 cell lysate
Lane 2 : ERBB2 knockout A549 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : SK-BR-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-ErbB2 / HER2 antibody [CAL27] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab237715 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type A549 cell lysates with no signal observed at this size in ERBB2 knockout cell line. To generate this image, wild-type and ERBB2 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237715).
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All lanes : Anti-ErbB2 / HER2 antibody [CAL27] (ab237715) at 1/1000 dilution
Lane 1 : Wild-type HCT 116 cell lysate
Lane 2 : ERBB2 knockout HCT 116 cell lysate
Lane 3 : HeLa cell lysate
Lane 4 : SK-BR-3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 137 kDa
Observed band size: 180 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-ErbB2 / HER2 antibody [CAL27] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab237715 was shown to bind specifically to ErbB2 / HER2. A band was observed at 180 kDa in wild-type HCT 116 cell lysates with no signal observed at this size in ERBB2 knockout cell line. To generate this image, wild-type and ERBB2 knockout HCT 116 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237715).
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Immunohistochemical analysis of human breast carcinoma tissue labeling ErbB2 / HER2 with ab237715 at 1/2000 dilution, followed by Rabbit specific IHC polymer detection kit HRP/DAB (ab209101). Positive staining on the human breast carcinoma is observed. Counter stained with hematoxylin.
Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Rabbit specific IHC polymer detection kit HRP/DAB (ab209101).
Heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0, epitope retrieval solution 2) for 20 mins.
The section was incubated with ab237715 for 10 mins at room temperature.
The immunostaining was performed on a Leica Biosystems BOND® RX instrument.
This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237715).
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Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilised SK-BR-3 (human mammary gland adenocarcinoma cell line) cells labeling ErbB2 / HER2 with ab237715 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing membranous staining in SK-BR-3 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) (ab195889) at 1/200 dilution (red).
Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (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 and sodium azide (ab237715).
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ErbB2 / HER2 was immunoprecipitated from 0.35 mg of HeLa (human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab237715 at 130 dilution. Western blot was performed from the immunoprecipitate using ab237715 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used as secondary antibody at 1/5000 dilution.
Lane 1: HeLa whole cell lysate 10 μg (Input).
Lane 2: ab237715 IP in HeLa whole lysate.
Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab237715 in HeLa whole cell lysate.Blocking and dilution buffer and concentration: 5% NFDM/TBST.
Exposure time: 15 seconds.This data was developed using the same antibody clone in a different buffer formulation containing PBS and sodium azide (ab237715).
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Intracellular flow cytometric analysis of4% paraformaldehyde-fixed, 90% methonao-permeabilized SH-BR-3 (human mammary gland adenocarcinoma cell line) cells labeling ErbB2 / HER2 with ab237715 at 1/500 dilution (red) compared with Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody) (blue).
Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077), 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 and sodium azide (ab237715).
Protocols
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
Datasheets and documents
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Datasheet download
References (0)
ab251602 has not yet been referenced specifically in any publications.