Overview

  • Product name
    Anti-ErbB 2 antibody [3B5]
    See all ErbB 2 primary antibodies
  • Description
    Mouse monoclonal [3B5] to ErbB 2
  • Tested applications
    Suitable for: IHC-Fr, IHC-P, WB, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human, Monkey
    Predicted to work with: Dog
  • Immunogen

    Synthetic peptide:

    TAENPEYLGLDVPV

    , corresponding to C terminal amino acids 1242 - 1255 of Human c-ErbB2.

  • Epitope
    Within amino acids 1242 - 1255 of human c-neu (TAENPEYLGLDVPV)
  • Positive control
    • SK-BR-3 cells or breast carcinoma tissue
  • General notes
    For paraffin sections, we recommend pretreating with a pressure cooker, but trypsin and heat will also work well. Including 0.05% saponin during staining of paraffin sections may help reduce background. In one study ductal carcinomas in situ displaying large-cell, comedo growth type, were stained while none of 16 ductal carcinomas in situ of small-cell, papillary, or cribriform growth type were stained by this antibody. MO 01/02/07: update consignment level to 0 following settlement of consignment stock in January 07.

Properties

Applications

Our Abpromise guarantee covers the use of ab16901 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
IHC-Fr Use a concentration of 2.5 µg/ml.
IHC-P Use at an assay dependent concentration.
WB Use a concentration of 2.5 µg/ml. Detects a band of approximately 190 kDa (predicted molecular weight: 138 kDa).
IP Use 1µg for 106 cells.
Flow Cyt 1/50 - 1/100.

ab170190-Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

Target

  • 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
    modifications
    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).
  • 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
  • Database links
  • Alternative names
    • Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog antibody
    • C erb B2/neu protein antibody
    • CD340 antibody
    • CD340 antigen antibody
    • Cerb B2/neu protein antibody
    • CerbB2 antibody
    • Erb b2 receptor tyrosine kinase 2 antibody
    • ERBB2 antibody
    • ERBB2_HUMAN antibody
    • HER 2 antibody
    • HER 2/NEU antibody
    • HER2 antibody
    • Herstatin antibody
    • Human epidermal growth factor receptor 2 antibody
    • Metastatic lymph node gene 19 protein antibody
    • MLN 19 antibody
    • MLN19 antibody
    • NEU antibody
    • NEU proto oncogene antibody
    • Neuro/glioblastoma derived oncogene homolog antibody
    • Neuroblastoma/glioblastoma derived oncogene homolog antibody
    • NGL antibody
    • p185erbB2 antibody
    • Proto-oncogene c-ErbB-2 antibody
    • Proto-oncogene Neu antibody
    • Receptor tyrosine-protein kinase erbB-2 antibody
    • TKR1 antibody
    • Tyrosine kinase type cell surface receptor HER2 antibody
    • Tyrosine kinase-type cell surface receptor HER2 antibody
    • V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 (neuro/glioblastoma derived oncogene homolog) antibody
    • V erb b2 avian erythroblastic leukemia viral oncogene homolog 2 antibody
    • V erb b2 avian erythroblastic leukemia viral oncoprotein 2 antibody
    • V erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian) antibody
    • V erb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog antibody
    • Verb b2 erythroblastic leukemia viral oncogene homolog 2, neuro/glioblastoma derived oncogene homolog (avian) antibody
    see all

Images

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast cancer tissue sections labeling ErbB 2 with ab16901 at 1/400 dilution. the tissue was fixed with formaldehyde; heat mediated antigen retrieval was performed using a acetate buffer pH6. An undiluted goat anti-mouse HRP conjugated seconday antibody was used.

    See Abreview

  • Anti-ErbB 2 antibody [3B5] (ab16901) at 1/250 dilution + Human Huh7 whole cell lysate at 20 µg

    Secondary
    Goat anti-mouse HRP conjugate at 1/5000 dilution

    Predicted band size : 138 kDa
    Observed band size : 138 kDa
    Additional bands at : 48 kDa (possible non-specific binding).

    Exposure time : 5 minutes

    This image is courtesy of an anonymous abreview.

    Blocking: 5% milk for 1 hour at 25°C.

    See Abreview

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human Mamma tumor Her2Neu tissue labeling ErbB 2 with ab16901.

  • Anti-ErbB 2 antibody [3B5] (ab16901) at 1 µg/ml + Whole cell lysate HEK293

    Predicted band size : 138 kDa
  • Overlay histogram showing LoVo cells stained with ab16901 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab16901, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in LoVo cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human breast carcinoma labeling ErbB 2 with ab16901.

References

This product has been referenced in:
  • Sweeney WE  et al. Tesevatinib ameliorates progression of polycystic kidney disease in rodent models of autosomal recessive polycystic kidney disease. World J Nephrol 6:188-200 (2017). Read more (PubMed: 28729967) »
  • Sui M  et al. Upregulation of miR-125b is associated with poor prognosis and trastuzumab resistance in HER2-positive gastric cancer. Exp Ther Med 14:657-663 (2017). Read more (PubMed: 28672982) »

See all 12 Publications for this product

Customer reviews and Q&As

Thank you for contacting us.

I am pleased to let you know that all our products are covered by our Abpromise (www.abcam.com/Abpromise):

We guarantee our products work in the tested species and applications as stated on the datashe...

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Breast cancer cell lines)
Permeabilization
Yes - 0.5% Triton-x/PBS
Specification
Breast cancer cell lines
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
Fixative
Paraformaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 03 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (Breast carcinoma and Breast cell lines)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate pH6 and EDTA pH8.5
Permeabilization
No
Specification
Breast carcinoma and Breast cell lines
Blocking step
(agent) for 10 minute(s) · Concentration: 50% · Temperature: 24°C
Fixative
Formaldehyde
Username

Mr. Malcolm Lim

Verified customer

Submitted Jun 27 2017

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Mammary gland from Her2-positive mouse)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Citrate ph 6
Permeabilization
Yes - Triton X-100 0.2% for 30 min
Specification
Mammary gland from Her2-positive mouse
Blocking step
Mouse IgG blocking reagent as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 90µg/mL · Temperature: 25°C
Fixative
Formaldehyde
Username

Flavia Ghiraldini

Verified customer

Submitted Apr 11 2017

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Dog Cell lysate - whole cell (MDCK)
Gel Running Conditions
Reduced Non-Denaturing (Native) (4-12%)
Loading amount
15 µg
Specification
MDCK
Blocking step
Milk as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Dec 19 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Flow Cytometry
Sample
Human Cell (N87 cell line)
Permeabilization
No
Gating Strategy
no gating
Specification
N87 cell line
Preparation
Cell harvesting/tissue preparation method: Cells scrapped in PBS
Sample buffer: PBS with 0.5% BSA
Fixation
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jul 06 2016

Application
Western blot
Sample
Human Cell lysate - whole cell (N87 cells)
Gel Running Conditions
Reduced Denaturing (10% Tris Glycin gel)
Loading amount
400000 cells
Specification
N87 cells
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jun 22 2016

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Human Tissue sections (human breast cancer)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: Acetate pH6
Permeabilization
No
Specification
human breast cancer
Blocking step
Innovex Background Buster as blocking agent for 30 minute(s) · Concentration: 100% · Temperature: 20°C
Fixative
Formaldehyde
Username

Abcam user community

Verified customer

Submitted Jun 15 2016

Application
Western blot
Loading amount
20 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (Huh7 (liver))
Specification
Huh7 (liver)
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 30 2014



It does sound like the buffer additives are interfering with the conjugation reaction. The gelatin and azide contain free amine groups that will interfere with this kind of reaction. I would suggest doing a buffer exchange or using a purifica...

Read More

1-10 of 19 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

Sign up