Product Pathways - Translational Control
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb (Alexa Fluor® 647 Conjugate) #4851
| Applications | Reactivity | Sensitivity | Source | Isotype |
|---|---|---|---|---|
| IF-IC IF-P F | H R (M) (Mk) | 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
Species enclosed in parentheses are predicted to react based on 100% sequence homology. Species cross-reactivity is determined by Western blot.
Specificity / Sensitivity
Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb detects endogenous levels of ribosomal protein S6 only when phosphorylated at Ser235 and 236. 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.
Source / Purification
Monoclonal antibodies are produced by immunizing rabbits with a synthetic phosphopeptide (KLH-coupled) corresponding to residues surrounding Ser235 and Ser236 of human ribosomal protein S6.
Flow Cytometry
Flow cytometric analysis of K562 cells, untreated (green) or STI571-treated (blue), using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) 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) Rabbit mAb (Alexa Fluor® 647 Conjugate).
IF-P
Confocal immunofluorescent analysis of paraffin-embedded H3255 xenograft using Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb (Alexa Fluor® 647 Conjugate) (blue) and Pan-Keratin (C11) Mouse mAb (Alexa Fluor® 488 Conjugate) #4523 (green).
Description
Cell Signaling Technology antibody is conjugated to Alexa Fluor® 647 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells. The unconjugated antibody #4858 reacts with human, monkey, mouse and rat Phospho-S6 Ribosomal protein. CST expects that Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb (Alexa Fluor® 647 Conjugate) will also recognize Phospho-S6 in these species.
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
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Companion Products
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- 4374 Phospho-p44/42 MAPK (Thr202/Tyr204) (E10) Mouse mAb (Alexa Fluor® 488 Conjugate)
- 3452 Rabbit IgG Isotype Control (Alexa Fluor® 647 Conjugate)
- 2317 S6 Ribosomal Protein (54D2) Mouse mAb
Alexa Fluor® is a registered trademark of Molecular Probes, Inc.
This product is for in vitro research use only and is not intended for use in humans or animals. This product is not intended for use as therapeutic or in diagnostic procedures.
