Cell Signaling Technology

Product Pathways - PathScan ELISA

PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Antibody Pair #7201

Kit Includes Volume Cap Color
S6 Ribosomal Protein Capture Antibody (100X) 0.4 milliliters Pink
Phospho-S6 Ribosomal Protein (Ser235/236) Detection Antibody (100X) 0.4 milliliters Blue
Anti-Rabbit IgG HRP-Linked Antibody (1000X) 0.04 milliliters Red

Capture and Detection Antibodies are stored at 4°C. HRP-Linked Secondary Antibody is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse

Description

CST's PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Antibody Pair is being offered as an alternative to our PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Kit #7205. Capture and Detection antibodies (100X stocks) and an HRP-conjugated secondary antibody (1000X stock) are supplied. Sufficient reagents are provided for performing 4 x 96 well ELISAs. The S6 Ribosomal Protein Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by Phospho-S6 Ribosomal Protein (Ser 235/236) Detection Antibody and HRP-conjugated secondary antibody. HRP substrate, TMB, is added for color development. The magnitude of the absorbance at 450 nm is proportional to the quantity of phospho-S6 Ribosomal protein (Ser235/236) protein.*Antibodies in this kit are custom formulations specific to the kit.

Sandwich ELISA

Sandwich ELISA

The relationship between lysate protein concentration from untreated and PDGF treated NIH/3T3 cells and the absorbance at 450 nm using PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Antibody Pair #7201 is shown. NIH/3T3 cells were starved overnight, treated with PDGF for 20 minutes at 37°C and then lysed.

Background

One way that growth factors and mitogens effectively promote sustained cell growth and proliferation is by upregulating mRNA translation (1,2). Growth factors and mitogens induce the activation of p70 S6 kinase and the subsequent phosphorylation of the S6 ribosomal protein. Phosphorylation of S6 ribosomal protein correlates with an increase in translation of mRNA transcripts that contain an oligopyrimidine tract in their 5' untranslated regions (2). These particular mRNA transcripts (5'TOP) encode proteins involved in cell cycle progression as well as ribosomal proteins and elongation factors necessary for translation (2,3). Important S6 ribosomal protein phosphorylation sites include several residues (Ser235, Ser236, Ser240 and Ser244) located within a small, carboxy-terminal region of the S6 protein (4,5).

  1. Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
  2. Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.
  3. Jefferies, H.B. et al. (1997) EMBO J. 16, 3693-3704.
  4. Ferrari, S. et al. (1991) J. Biol. Chem. 266, 22770-22775.
  5. Flotow, H. and Thomas, G. (1992) J. Biol. Chem. 267, 3074-3078.

Application References

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