Clone:
REA152
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
MAPK1, MAPK3, ERK, ERK-2, ERK-1

Extended validation for ERK1/2 pT202/pY204 Antibody, anti-human/mouse, 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 REA152
20A++
MILAN8R+
4B11B69 (mIgG2b)n/a
6B8B69++
ERK12T202Y204-A11n/a
4B11B69++
Cells were incubated with an excess of purified unconjugated ERK1/2 pT202/pY204 (REA152) 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 ERK1/2 pT202/pY204. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 40nM PMA for 10 min at 37°C. Cells were stained with ERK1/2 pT202/pY204 antibodies and plotted against the side scatter. As a control, ERK1/2 pT202/pY204 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, 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 ERK1/2 pT202/pY204. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 40nM PMA for 10 min at 37°C. Cells were stained with ERK1/2 pT202/pY204 antibodies and plotted against the side scatter. As a control, ERK1/2 pT202/pY204 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, 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 ERK1/2 pT202/pY204. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 40nM PMA for 10 min at 37°C. Cells were stained with ERK1/2 pT202/pY204 antibodies and plotted against the side scatter. As a control, ERK1/2 pT202/pY204 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, 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 ERK1/2 pT202/pY204. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 40nM PMA for 10 min at 37°C. Cells were stained with ERK1/2 pT202/pY204 antibodies and plotted against the side scatter. As a control, ERK1/2 pT202/pY204 antibody staining was omitted and cells were measured in the same channels. Flow cytometry was performed with the MACSQuant® Analyzer. Cell debris, dead cells, 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 ERK1/2 pT202/pY204 Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA152 recognizes the phosphorylated threonine 202 and tyrosine 204 (pT202/pY204) of human ERK1 and pT184/pY186 of human ERK2, which are key components of the MAPK signaling cascade. ERK1 and ERK2 are multifunctional serine/threonine kinases and are ubiquitously expressed in mammalian cells. They share 85% homology and are phosphorylated by kinases MEK1/2 at Thr and Tyr residues within their activation loop leading to activation and nuclear translocation of ERKs. Activated ERKs phosphorylate downstream targets, such as signaling effectors, receptors, cytoskeletal proteins, and transcription factors. Owing to such a wide array of cellular targets, ERKs are involved in regulation of a number of processes including cell adhesion, cell cycle progression, cell migration, cell survival, differentiation, metabolism, proliferation, and transcription. Activation and mobilization of ERKs is induced in response to several extracellular signals and engagement of receptors such as Tyr kinases (RTKs), G protein–coupled receptors (GPCRs), ion channels, and others. The ERK1/2 are dual specificity kinases and phosphorylate Ser or Thr residues neighbor to Proline residue.
Additional information: Clone REA152 displays negligible binding to Fc receptors.

Alternative names

MAPK1, MAPK3, ERK, ERK-2, ERK-1

Detailed product information

Technical specifications

CloneREA152
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse, rat
Cross-reactivityrat
AntigenERK1/2 pT202/pY204
Alternative names of antigenMAPK1, MAPK3, ERK, ERK-2, ERK-1
Molecular mass of antigen [kDa]42
Distribution of antigenother
Entrez Gene ID5595, 5594
RRIDAB_2651665, AB_2651666, AB_2651667, AB_2651668, AB_2651669, AB_2651664

Resources for ERK1/2 pT202/pY204 Antibody, anti-human/mouse, REAfinity™

Certificates

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References for ERK1/2 pT202/pY204 Antibody, anti-human/mouse, REAfinity™

Publications

  1. Davis, R. J. et al. (1995) Transcriptional regulation by MAP kinases. Mol. Reprod. Dev. 42(4): 459-467
  2. Roskoski, R. et al. (2012) ERK1/2 MAP kinases: structure, function, and regulation. Pharmacol. Res. 66(2): 105-143
  3. Lloyd, A. C. (2006) Distinct functions for ERKs? J. Biol. 5(5): 13
  4. Gonzalez-Perez, G. et al. (2017)
    Gastrointestinal microbiome dysbiosis in infant mice alters peripheral CD8
    +
    T cell receptor signaling.
    Front Immunol 8: 265

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