Clone:
REA420
Type of antibody:
Primary antibodies, Recombinant antibodies
Isotype:
recombinant human IgG1
Applications:
ICFC
Alternative names:
RPS6, NP33, RS6

Extended validation for S6 pS240 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 REA420
N4-41++
Cells were incubated with an excess of purified unconjugated S6 pS240 (REA420) 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 pS240. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 200ng/ml Phorol-12-myristat-13-acetat (PMA) for 30min at 37°C. Cells were fixed and permeabiliz using the Cell Signaling Buffer Set A and stained with S6 pS240 antibodies and plotted against the side scatter. As a control, S6 pS240 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 S6 pS240. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 200ng/ml Phorol-12-myristat-13-acetat (PMA) for 30min at 37°C. Cells were fixed and permeabiliz using the Cell Signaling Buffer Set A and stained with S6 pS240 antibodies and plotted against the side scatter. As a control, S6 pS240 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 S6 pS240. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 200ng/ml Phorol-12-myristat-13-acetat (PMA) for 30min at 37°C. Cells were fixed and permeabiliz using the Cell Signaling Buffer Set A and stained with S6 pS240 antibodies and plotted against the side scatter. As a control, S6 pS240 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 S6 pS240. Human peripheral blood mononuclear cells (PBMCs) were stimulated with 200ng/ml Phorol-12-myristat-13-acetat (PMA) for 30min at 37°C. Cells were fixed and permeabiliz using the Cell Signaling Buffer Set A and stained with S6 pS240 antibodies and plotted against the side scatter. As a control, S6 pS240 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 S6 pS240 Antibody, anti-human, REAfinity™

Overview

Clone REA420 recognizes the human 40S ribosomal protein S6 antigen phosphorylated at serine 240 (pS240). 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.
Additional information: Clone REA420 displays negligible binding to Fc receptors.

Alternative names

RPS6, NP33, RS6

Detailed product information

Technical specifications

CloneREA420
Clonalitymonoclonal
Isotyperecombinant human IgG1
Isotype controlREA Control Antibody (I), human IgG1
Hostcell line
Type of antibodyPrimary antibodies, Recombinant antibodies
Specieshuman
AntigenS6 pS240
Alternative names of antigenRPS6, NP33, RS6
Molecular mass of antigen [kDa]29
Entrez Gene ID6194
RRIDAB_2857418, AB_2653390, AB_2653391, AB_2653392, AB_2653393, AB_2857419

Resources for S6 pS240 Antibody, anti-human, REAfinity™

Certificates

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References for S6 pS240 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. Shah, O. J. et al. (2003) Mitotic regulation of ribosomal S6 kinase 1 involves Ser/Thr, Pro phosphorylation of consensus and non-consensus sites by Cdc2. J. Biol. Chem. 278(18): 16433-16442

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