Product Pathways - Translational Control
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #4851
PhosphoSitePlus® protein, site, and accession data: S6
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| IF-IC IF-P F | H M R Mk Sc | Endogenous | Rabbit IgG |
Applications Key: IF-IC=Immunofluorescence (Immunocytochemistry) IF-P=Immunofluorescence (Paraffin) F=Flow Cytometry
Reactivity Key: H=Human M=Mouse R=Rat Mk=Monkey Sc=S. cerevisiae
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.
Protocols
- 4851:
- Flow, Immunofluorescence
Specificity / Sensitivity
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) detects endogenous levels of ribosomal protein S6 only when phosphorylated at Ser235 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. The antibody was conjugated to Alexa Fluor® 647 under optimal conditions with an F/P ratio of 2-5. The Alexa Fluor® 647 dye is maximally excited by red light (e.g. 633 nm He-Ne laser). Antibody conjugates of the Alexa Fluor® 647 dye produce bright far-red-fluorescence emission, with a peak at 665 nm.
Flow Cytometry
Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue), using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate).
IF-IC
Confocal immunofluorescent analysis of HeLa cells treated with 20% serum (left) or with Rapamycin, Wortmannin and U0126 inhibitors (right) using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) #4851.
IF-P
Confocal immunofluorescent analysis of paraffin-embedded H3255 xenograft using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue pseudocolor) and Pan-Keratin (C11) Mouse mAb (Alexa Fluor® 488 Conjugate) #4523 (green).
Description
This Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometry and immunofluorescent analysis in human cells. The antibody is expected to exhibit the same species cross-reactivity as the unconjugated Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb #4858.
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).
- Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
- Peterson, R.T. and Schreiber, S.L. (1998) Curr. Biol. 8, R248-R250.
- Jefferies, H.B. et al. (1997) EMBO J. 16, 3693-3704.
- Ferrari, S. et al. (1991) J. Biol. Chem. 266, 22770-22775.
- Flotow, H. and Thomas, G. (1992) J. Biol. Chem. 267, 3074-3078.
Application References
- Soond, D.R. et al. (2010) Blood 115, 2203-13. Applications: Flow Cytometry
- Francisco, L.M. et al. (2009) J Exp Med 206, 3015-29. Applications: Flow Cytometry
- Kalland, M.E. et al. (2011) J Immunol 187, 5233-45. Applications: Flow Cytometry
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Companion Products
- 4803 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate)
- 4858 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb
- 2336 Phospho-Akt (Ser473) (193H12) Rabbit mAb (Alexa Fluor® 488 Conjugate)
- 4374 Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 488 Conjugate)
- 3452 Rabbit IgG Isotype Control (Alexa Fluor® 647 Conjugate)
- 2217 S6 Ribosomal Protein (5G10) Rabbit mAb
- 2211 Phospho-S6 Ribosomal Protein (Ser235/236) Antibody
- 4856 Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb
- 4857 Phospho-S6 Ribosomal Protein (Ser235/236) (91B2) Rabbit mAb
- 2215 Phospho-S6 Ribosomal Protein (Ser240/244) Antibody
- 2985 Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 647 Conjugate)
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
For Research Use Only. Not For Use In Diagnostic Procedures.
