Cell Signaling Technology

Product Pathways - Translational Control

Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb #4856

Applications Reactivity Sensitivity MW (kDa) Isotype
W IF-IC F H M R Mk Endogenous 32 Rabbit IgG

Applications Key:  W=Western Blotting  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb detects endogenous levels of ribosomal protein S6 only when phosphorylated at serine 235 and 236.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic phosphopeptide corresponding to residues surrounding Ser235 and Ser236 of human ribosomal protein S6.

Western Blotting

Western Blotting

Western blot analysis of extracts from NIH/3T3 cells, untreated or PDGF-treated (100 ng/ml, 20 min), using Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (upper) or S6 Ribosomal Protein (5G10) Rabbit mAb #2217 (lower).

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of Jurkat cells, untreated (green), or LY294002, Wortmannin and U0126-treated (blue), using Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb compared to a nonspecific negative control antibody (red).

IF-IC

IF-IC

Confocal immunofluorescent images of HeLa cells serum-starved for 20 hrs (left), 20% serum-treated (center), or 20% serum-treated after preincubation with Rapamycin (FRAP/mTOR Inhibitor) #9904 and labeled with Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (green). Actin filaments have been labeled with Alexa Fluor® 555 phalloidin (red). Blue pseudocolor = DRAQ5™ (fluorescent DNA dye).


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.
  6. Kotecha, N. et al. (2008) Cancer Cell 14, 335-43.

Application References

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Companion Products

Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.

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

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