Clone:
REA375
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
MAP2K1, CFC3, MAPKK 1, MKK1, PRKMK1, MEK1

Extended validation for MEK1 pS298 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 REA375
J114-64++
MEK1S298-H8-
A16117B-
Cells were incubated with an excess of purified unconjugated MEK1 pS298 (REA375) 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 MEK1 pS298. A epithelial cell (HeLa) cells were stimulated with 150nM calyculin A for 30min. Cells were then fixed and permeabilized using the Cell signalling buffer set A MEK1 pS298 antibodies and plotted against the side scatter. As a control, MEK1 pS298 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 MEK1 pS298. A epithelial cell (HeLa) cells were stimulated with 150nM calyculin A for 30min. Cells were then fixed and permeabilized using the Cell signalling buffer set A MEK1 pS298 antibodies and plotted against the side scatter. As a control, MEK1 pS298 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 MEK1 pS298. A epithelial cell (HeLa) cells were stimulated with 150nM calyculin A for 30min. Cells were then fixed and permeabilized using the Cell signalling buffer set A MEK1 pS298 antibodies and plotted against the side scatter. As a control, MEK1 pS298 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 MEK1 pS298. A epithelial cell (HeLa) cells were stimulated with 150nM calyculin A for 30min. Cells were then fixed and permeabilized using the Cell signalling buffer set A MEK1 pS298 antibodies and plotted against the side scatter. As a control, MEK1 pS298 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 MEK1 pS298 Antibody, anti-human/mouse, REAfinity™

Overview

Clone REA375 recognizes the human and mouse MEK1 antigen phosphorylated at serine 298 (pS298). The dual specificity kinases MEK1 and MEK2 are activated by RAF and mediate phosphorylation of ERK1 and ERK2. MEK1 and MEK2 are very similar but differ structurally in a proline-rich domain in the C-terminal half of the catalytic core. Binding of extracellular ligands such as growth factors, cytokines and hormones to their cell-surface receptors activates RAS and this initiates RAF1 activation. Depending on the cellular context, this pathway mediates diverse biological functions such as cell growth, adhesion, survival, and differentiation, predominantly through the regulation of transcription, metabolism, and cytoskeletal rearrangements. MEK1 is phosphorylated at S298 by PAK. MAPK1/ERK2 phosphorylation of T292 occurs in response to cellular adhesion and leads to inhibition of S298 phosphorylation by PAK.
Additional information: Clone REA375 displays negligible binding to Fc receptors.

Alternative names

MAP2K1, CFC3, MAPKK 1, MKK1, PRKMK1, MEK1

Detailed product information

Technical specifications

CloneREA375
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman, mouse
AntigenMEK1 pS298
Alternative names of antigenMAP2K1, CFC3, MAPKK 1, MKK1, PRKMK1, MEK1
Molecular mass of antigen [kDa]43
Entrez Gene ID5604
RRIDAB_2652846, AB_2652847

Resources for MEK1 pS298 Antibody, anti-human/mouse, REAfinity™

Certificates

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References for MEK1 pS298 Antibody, anti-human/mouse, REAfinity™

Publications

  1. Zheng, C. F. et al. (1994) Activation of MEK family kinases requires phosphorylation of two conserved Ser/Thr residues. EMBO J. 13(5): 1123-1131
  2. Zheng, C. F. et al. (1993) Cloning and characterization of two distinct human extracellular signal-regulated kinase activator kinases, MEK1 and MEK2. J. Biol. Chem. 268(15): 11435-11439
  3. Coles, L. C. et al. (2002) PAK1 primes MEK1 for phosphorylation by Raf-1 kinase during cross-cascade activation of the ERK pathway. Oncogene 21(14): 2236-2244

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