Product Pathways - Translational Control
Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854
| Applications | Reactivity | Sensitivity | Isotype |
|---|---|---|---|
| IF-F IF-IC F | H R | Endogenous | Rabbit IgG |
Applications Key:
IF-F=Immunofluorescence (Frozen)
IF-IC=Immunofluorescence (Immunocytochemistry)
F=Flow Cytometry
Reactivity Key:
H=Human
R=Rat
Species cross-reactivity is determined by Western blot.
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 antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Ser235 and Ser236 of human ribosomal protein S6. The antibody was conjugated to Alexa Fluor® 488 under optimal conditions with an F/P ratio of 2-6.
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 (Alexa Fluor® 488 Conjugate).
IF-IC
Confocal immunofluorescent analysis of two Gefinitib (Iressa) treated non-small cell lung cancer cell lines. HCC827 cells have the E746_A750 deletion in exon 19 of the EGFR gene, and are highly sensitive to Gefitinib. H1975 cells have the (T790M) mutation that confers Gefitinib-resistance. Both cell lines were treated and then double-labeled with Phospho-S6 Ribosomal Protein Rabbit mAb (Alexa Fluor® 488 Conjugate) #4854 and Phospho-Tyrosine Mouse mAb (Alexa Fluor® 647 Conjugate) #9415. Untreated HCC827 (A) and H1975 (C) cells show bright phospho-S6 (green) and phospho-tyrosine (red pseudocolor) label. Phospho-S6 and phospho-tyrosine signals dramatically decrease following Gefitinib treatment in HCC827 cells (B), with little or no change in the Gefitinib-resistant H1975 cells (D).
IF-F
Confocal immunofluorescent analysis of rat brain showing the hippocampus (left) and striatum (right) labeled with Phospho-S6 Ribosomal Protein (Ser235/236) (2F9) Rabbit mAb (Alexa Fluor® 488 Conjugate) (green), EGR1 Antibody (red) #4152, and Phospho-Histone H3 (Ser10) (6G3) Mouse mAb #9706 (blue) following ischemia with 30 minute reperfusion.
Description
Cell Signaling Technology Antibody conjugated to Alexa Fluor® 488 fluorescent dye and tested in-house for direct flow cytometric analysis of human cells.*The unconjugated antibody, #4856, reacts with human and mouse phospho-S6 ribosomal protein. CST expects that Phospho-S6 Ribosomal Protein (2F9) Rabbit mAb (Alexa Fluor® 488 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
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
- 4858 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb
- 4851 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb (Alexa Fluor® 647 Conjugate)
- 4803 Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) Rabbit mAb (Alexa Fluor® 488 Conjugate)
Rabbit Monoclonals Produced Using Epitomics® Technology, U.S. Patent No. 5,675,063.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.
