Cell Signaling Technology

Product Pathways - PathScan ELISA

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

Kit Includes Volume Cap Color
P-S6 Ribosomal Protein (S235/236) Capture Ab (100X) 0.4 ml Pink
S6 Ribosomal Protein Detection Ab (100X) 0.4 ml Blue
Anti-mouse IgG, HRP-linked Antibody (1000X) 0.04 ml Red

Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.

Species Cross-Reactivity

H M

Reactivity Key:  H=Human  M=Mouse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Description

CST's PathScan® Phospho-S6 Ribosomal Protein (Ser235/236) Sandwich ELISA Antibody Pair is 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 Phospho-S6 Ribosomal Protein (Ser235/236) Capture Antibody is coated in PBS overnight in a 96 well microplate. After blocking, cell lysates are added, followed by S6 Ribosomal Protein 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 kit are custom formulations specific to kit.

Specificity / Sensitivity

For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.

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 37ºC 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

Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!

Companion Products

For Research Use Only. Not For Use In Diagnostic Procedures.

Products