Clone:
REA454
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
RPS6, NP33, RS6

Extended validation for S6 pS235/pS236 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 REA454
N7-548++
cupk43k+
A17020B++
Cells were incubated with an excess of purified unconjugated S6 pS235/pS236 (REA454) 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 S6 pS235/pS236. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA for 15min followed by a fixation and permeabilization step using the Cell Signaling Buffer Set A. Cells were then stained with S6 pS235/pS236 antibodies and plotted against the side scatter. As a control, S6 pS235/pS236 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 S6 pS235/pS236. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA for 15min followed by a fixation and permeabilization step using the Cell Signaling Buffer Set A. Cells were then stained with S6 pS235/pS236 antibodies and plotted against the side scatter. As a control, S6 pS235/pS236 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 S6 pS235/pS236. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA for 15min followed by a fixation and permeabilization step using the Cell Signaling Buffer Set A. Cells were then stained with S6 pS235/pS236 antibodies and plotted against the side scatter. As a control, S6 pS235/pS236 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 S6 pS235/pS236. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA for 15min followed by a fixation and permeabilization step using the Cell Signaling Buffer Set A. Cells were then stained with S6 pS235/pS236 antibodies and plotted against the side scatter. As a control, S6 pS235/pS236 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 S6 pS235/pS236. Human peripheral blood mononuclear cells (PBMCs) were stimulated with PMA for 15min followed by a fixation and permeabilization step using the Cell Signaling Buffer Set A. Cells were then stained with S6 pS235/pS236 antibodies and plotted against the side scatter. As a control, S6 pS235/pS236 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 S6 pS235/pS236 Antibody, anti-human, REAfinity™

Overview

Clone REA454 recognizes the human 40S ribosomal protein S6 antigen phosphorylated at serine 235 and 236 (pS235/pS236). Ribosomal protein S6 is one of 33 proteins that comprise the 40 S ribosomal subunit and becomes phosphorylated at several serine residues upon mitogen stimulation. It plays an important role in controlling cell growth and proliferation through the selective translation of particular classes of mRNA. Activation of the p90 ribosomal S6 kinases (RSKs) by serum, growth factors, tumor promoting phorbol esters, and oncogenic Ras is required for ribosomal protein S6 phosphorylation downstream of the Ras/ERK signaling cascade. The phosphorylation sites in ribosomal protein S6 have been mapped to five clustered serine residues – S235, S236, S240, S244, and S247 – which are located at the C terminus and are conserved from Drosophila to mammals. While ribosomal S6 kinase 1 phosphorylates ribosomal protein S6 at all sites, RSK exclusively phosphorylates ribosomal protein S6 at S235 and S236
in vitro
and
in vivo
using an mTOR-independent mechanism.
Additional information: Clone REA454 displays negligible binding to Fc receptors.

Alternative names

RPS6, NP33, RS6

Detailed product information

Technical specifications

CloneREA454
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenS6 pS235/pS236
Alternative names of antigenRPS6, NP33, RS6
Molecular mass of antigen [kDa]29
Entrez Gene ID6194

Resources for S6 pS235/pS236 Antibody, anti-human, REAfinity™

Certificates

Please follow this
link
to search for Certificates of Analysis (CoA) by lot number.

References for S6 pS235/pS236 Antibody, anti-human, REAfinity™

Publications

  1. Selman, C. et al. (2009) Ribosomal protein S6 kinase 1 signaling regulates mammalian life span. Science 326(5949): 140-144
  2. Lott, J. B. et al. (1988) Isolation and characterization of cloned cDNAs that code for human ribosomal protein S6. Gene 65(1): 31-39
  3. Roux, P. P. et al. (2007) RAS/ERK signaling promotes site-specific ribosomal protein S6 phosphorylation via RSK and stimulates cap-dependent translation. J. Biol. Chem. 282(19): 14056-14064

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