Clone:
REA439
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
EGFR, ERBB, HER1, NISBD2, PIG61, mENA, erbB-1

Extended validation for EGF Receptor pY845 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 REA439
12A3++
Cells were incubated with an excess of purified unconjugated EGF Receptor pY845 (REA439) 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
Flow cytometric comparison of different clones for EGF Receptor pY845. A-431 cells were stimulates for 24 hours serum starved and stimulated with 100 ng/mL human EGF for 30 minutes at 4 °C followed by incubation for 15 minutes at 37 °. Cells were stained with a suitable counterstaining followed by a fixation and permeabilization step using Cell Signalling buffer set. Cells were then stained with EGF Receptor pY845 antibodies and plotted against the side scatter. As a control, EGF Receptor pY845 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for EGF Receptor pY845. A-431 cells were stimulates for 24 hours serum starved and stimulated with 100 ng/mL human EGF for 30 minutes at 4 °C followed by incubation for 15 minutes at 37 °. Cells were stained with a suitable counterstaining followed by a fixation and permeabilization step using Cell Signalling buffer set. Cells were then stained with EGF Receptor pY845 antibodies and plotted against the side scatter. As a control, EGF Receptor pY845 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
View details
Flow cytometric comparison of different clones for EGF Receptor pY845. A-431 cells were stimulates for 24 hours serum starved and stimulated with 100 ng/mL human EGF for 30 minutes at 4 °C followed by incubation for 15 minutes at 37 °. Cells were stained with a suitable counterstaining followed by a fixation and permeabilization step using Cell Signalling buffer set. Cells were then stained with EGF Receptor pY845 antibodies and plotted against the side scatter. As a control, EGF Receptor pY845 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.
Flow cytometric comparison of different clones for EGF Receptor pY845. A-431 cells were stimulates for 24 hours serum starved and stimulated with 100 ng/mL human EGF for 30 minutes at 4 °C followed by incubation for 15 minutes at 37 °. Cells were stained with a suitable counterstaining followed by a fixation and permeabilization step using Cell Signalling buffer set. Cells were then stained with EGF Receptor pY845 antibodies and plotted against the side scatter. As a control, EGF Receptor pY845 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris and cell doublets were excluded from the analysis based on scatter signals. No FcR Blocking Reagent was used. The recommended titers of respective antibodies from different suppliers were used.

Specifications for EGF Receptor pY845 Antibody, anti-human, REAfinity™

Overview

Clone REA439 recognizes the human epidermal growth factor receptor (EGFR) antigen phosphorylated at tyrosine 845 (pY845). EGFR is a single-pass type I membrane protein which is also known as receptor tyrosine-protein kinase ErbB-1 or HER1. It is a critical regulator of many normal cellular processes, including cell growth, differentiation, survival, and migration. The EGFR also is implicated in pathological processes of cellular transformation and oncogenesis due to its overexpression in many types of cancers and its ability to induce morphological transformation of cultured cells and tumor formation in nude mice. Many different signaling pathways have been discovered to mediate the effects of EGF, the most notable being Ras-mitogen-activated protein kinase (MAPK) and the phosphatidylinositol 3-kinase pathways. In these pathways EGF binds to the EGFR to induce dimerization, catalytic activation, and autophosphorylation of tyrosines in the C-terminal tail of the EGFR. These phosphorylated tyrosines provide docking sites for adapter proteins such as Grb2, Shc, and Gab2 to link the receptor to the Ras-MAPK, as well as phosphatidylinositol 3-kinase pathways. Phosphorylation of the highly conserved tyrosine 845 is mediated by c-Src and dependent on EGF stimulation. Signals emanating from phosphorylated Y845 on the EGFR may involve effector molecules other than Shc and MAPK.
Additional information: Clone REA439 displays negligible binding to Fc receptors.

Alternative names

EGFR, ERBB, HER1, NISBD2, PIG61, mENA, erbB-1

Detailed product information

Technical specifications

CloneREA439
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hosthuman cell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse, rat
Cross-reactivitymouse, rat
AntigenEGF Receptor pY845
Alternative names of antigenEGFR, ERBB, HER1, NISBD2, PIG61, mENA, erbB-1
Molecular mass of antigen [kDa]132(refers to canonical form)
Distribution of antigenother
Entrez Gene ID1956
RRIDAB_2651614, AB_2651615, AB_2651616, AB_2651617

Resources for EGF Receptor pY845 Antibody, anti-human, REAfinity™

Certificates

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References for EGF Receptor pY845 Antibody, anti-human, REAfinity™

Publications

  1. Ullrich, A. et al. (1984) Human epidermal growth factor receptor cDNA sequence and aberrant expression of the amplified gene in A431 epidermoid carcinoma cells. Nature 309(5967): 418-425
  2. Heisermann, G. J. et al. (1988)
    Epidermal growth factor receptor threonine and serine residues phosphorylated
    in vivo
    .
    J. Biol. Chem. 263(26): 13152-13158
  3. Boerner, J. L. et al. (2004) Phosphorylation of Y845 on the epidermal growth factor receptor mediates binding to the mitochondrial protein cytochrome c oxidase subunit II. Mol. Cell. Biol. 24(16): 7059-7071

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