Clone:
REA1232
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
FC, MICS, IF, IHC
Alternative names:
p185erbB2, HER2, MLN19, NEU, NGL, HER2/neu

Extended validation for ErbB-2 (CD340) Antibody, anti-human, REAfinity™

Specificity

Epitope competition
In order to compare the epitope specificity of an antibody, the clone being used is compared with other known clones recognizing the same antigen in a competition assay.
Other clonesOverlap in epitope recognition with REA1232
24D2++
NEU.24.7-
2G11-
Cells were incubated with an excess of purified unconjugated ErbB-2 (CD340) (REA1232) antibody followed by staining with fluorochrome-conjugated antibodies of other known clones against the same marker. Based on the fluorescence signal obtained, the clones were identified as recognizing completely overlapping (++), partially overlapping (+), or completely different epitopes (-) of the marker.

Sensitivity

Performance comparison
Selected fluorochrome conjugated antibodies from Miltenyi Biotec were compared to commercially available hybridoma clones in flow cytometry analysis.
View details
MCF-7 cells were stained with Anti-ErbB-2 (CD340) antibodies or with the corresponding REA Control (S) antibodies (left peak). Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
View details
MCF-7 cells were stained with Anti-ErbB-2 (CD340) antibodies or with the corresponding REA Control (S) antibodies (left peak). Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
View details
MCF-7 cells were stained with Anti-ErbB-2 (CD340) antibodies or with the corresponding REA Control (S) antibodies (left peak). Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
View details
MCF-7 cells were stained with Anti-ErbB-2 (CD340) antibodies or with the corresponding REA Control (S) antibodies (left peak). Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
MCF-7 cells were stained with Anti-ErbB-2 (CD340) antibodies or with the corresponding REA Control (S) antibodies (left peak). Flow cytometry was performed with the MACSQuant® Analyzer. The Tandem Signal Enhancer has been used to increase binding specificity of tandem-dye–conjugated antibodies. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence or 4',6-diamidino-2-phenylindole (DAPI) fluorescence, as in the case of tandem conjugates.
Fixation data
To provide an indication on how an antibody performs after fixation of cells, in-house data on staining results before and after fixation with 3.7% formaldehyde using Miltenyi Biotec antibodies are provided. Different experimental settings may lead to different results.
No fixation
Post fixation
View details
Comparison of staining pattern on non-fixed and fixed cells using ErbB-2 (CD340) (REA1232). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
View details
Comparison of staining pattern on non-fixed and fixed cells using ErbB-2 (CD340) (REA1232). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.
Comparison of staining pattern on non-fixed and fixed cells using ErbB-2 (CD340) (REA1232). The performance of the antibody after fixation was tested by comparing the staining pattern on fresh (no fixation) versus fixed (post fixation) cells.

Specifications for ErbB-2 (CD340) Antibody, anti-human, REAfinity™

Overview

Clone REA1232 recognizes the human receptor tyrosine-protein kinase ErbB-2 antigen, a Single-pass type I membrane protein which is also known as CD340, human epidermal growth factor receptor 2 (Her2) or proto-oncogene Neu. ErbB-2 is a member of the epidermal growth factor receptor family. The ErbB family is composed of four plasma membrane-bound receptor tyrosine kinases. All four contain an extracellular ligand binding domain, a transmembrane domain, and an intracellular domain that can interact with a multitude of signaling molecules and exhibit both ligand-dependent and ligand-independent activity. ErbB-2 can heterodimerise with any of the other three receptors and is considered to be the preferred dimerisation partner of the other ErbB receptors. Dimerisation results in the autophosphorylation of tyrosine residues within the cytoplasmic domain of the receptors and initiates a variety of signaling pathways. ErbB-2 has been shown to be involved in embryonic development and cancer progression. Amplification or overexpression of this oncogene has been shown to play an important role in the development and progression of certain aggressive types of cancer. In recent years the protein has become an important biomarker and target of therapy for approx. 30% of breast cancer patients.
Additional information: Clone REA1232 displays negligible binding to Fc receptors.

Alternative names

p185erbB2, HER2, MLN19, NEU, NGL, HER2/neu

Detailed product information

Technical specifications

CloneREA1232
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (S), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenErbB-2 (CD340)
Alternative names of antigenp185erbB2, HER2, MLN19, NEU, NGL, HER2/neu
Molecular mass of antigen [kDa]136
Distribution of antigentumor cells
Entrez Gene ID2064
RRIDAB_2819625, AB_2819633, AB_2819626, AB_2819634, AB_2819627, AB_2819635, AB_2819628, AB_2819636, AB_2819629, AB_2819637, AB_2819630, AB_2819638, AB_2819631, AB_2819632

Resources for ErbB-2 (CD340) Antibody, anti-human, REAfinity™

Certificates

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References for ErbB-2 (CD340) Antibody, anti-human, REAfinity™

Publications

  1. Yamamoto, T. et al. (1986) Similarity of protein encoded by the human c-erb-B-2 gene to epidermal growth factor receptor. Nature 319(6050): 230-234
  2. Wang, Y. N. et al. (2012) Membrane-bound trafficking regulates nuclear transport of integral epidermal growth factor receptor (EGFR) and ErbB-2. J. Biol. Chem. 287(20): 16869-16879
  3. Tsutsui, S. et al. (2002) Prognostic value of c-erbB2 expression in breast cancer. J Surg Oncol 79(4): 216-223

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