Cell Signaling Technology

Product Pathways - Translational Control

Phospho-S6 Ribosomal Protein (Ser235/236) Blocking Peptide #1220

Description

This peptide is used to block Phospho-S6 Ribosomal Protein (Ser235/236) (91B2) Rabbit mAb #4857, Phospho-S6 Ribosomal Protein (Ser235/236) Antibody #2211, Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) (XP) Rabbit mAb #4858, and Phospho-S6 Ribosomal Protein (Ser235/236) (M20B19) Mouse mAb #5006 reactivity.

Quality Control

The quality of the peptide was evaluated by reversed-phase HPLC and by mass spectrometry. The peptide blocks Phospho-S6 Ribosomal Protein (Ser235/236) (91B2) Rabbit mAb #4857, Phospho-S6 Ribosomal Protein (Ser235/236) Antibody #2211, Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) (XP) Rabbit mAb #4858, and Phospho-S6 Ribosomal Protein (Ser235/236) (M20B19) Mouse mAb #5006 signal in immunohistochemistry and Western Blotting.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, using Phospho-S6 Ribosomal Protein (Ser235/236) (91B2) Rabbit mAb #4857 in the presence of control peptide (left) or Phospho-S6 Ribosomal Protein (Ser235/236) Blocking Peptide (right).

Western blot analysis of extracts from 3T3 cells, untreated or treated with 20% serum, using Phospho-S6 Ribosomal Protein (Ser235/236) Antibody #2211 (left), or the same antibody incubated with specific blocking peptide (right).

Applications

Use as a blocking reagent to evaluate the specificity of antibody reactivity in immunohistochemistry protocols.

Directions for Use

For immunohistochemistry, add twice the volume of peptide as volume of antibody used in 100 µl total volume. Incubate for a minimum of 30 minutes prior to adding the entire volume to the slide. Recommended antibody dilutions can be found on the product data sheet.

For Western immunoblotting, add 10 µl of antibody and 10 µl of blocking peptide to 10 ml of antibody dilution buffer, and incubate at room

temperature for 30 minutes before allowing to react with the blot.

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).

No product specific background information is currently available for this product.

  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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Companion Products

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

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