Product Pathways - Translational Control
PhosphoPlus® p70 S6 Kinase (Thr389, Thr421/Ser424) Antibody Kit #9430
| Kit Includes | Quantity | Applications | Reactivity | MW (kDa) | Source |
|---|---|---|---|---|---|
| Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb # 9234 | 100 microliters | W | H M R | 70, 85 | Rabbit |
| Phospho-p70 S6 Kinase (Thr421/Ser424) Antibody # 9204 | 100 microliters | W IP | H M R Mk Pg | 70, 85 | Rabbit |
| p70 S6 Kinase Antibody # 9202 | 100 microliters | W IP | H M R Mk | 70, 85 | Rabbit |
| Anti-rabbit IgG, HRP-linked Antibody # 7074 | 50 microliters | Goat | |||
| Anti-biotin, HRP-linked Antibody # 7075 | 100 microliters | Goat | |||
| Biotinylated Protein Ladder Detection Pack # 7727 | 100 microliters | ||||
| 20X LumiGLO® Reagent and 20X Peroxide # 7003 | 5 milliliters | ||||
| p70 S6 Kinase Control Cell Extracts # 9203 | 80 microliters |
Applications Key:
Reactivity Key:
Specificity / Sensitivity
p70 S6 Kinase Antibody #9202 detects endogenous levels of total p70 S6 Kinase. It also recognizes p85 S6 Kinase. Phospho-p70 S6 Kinase (Thr389) Antibody and Phospho-p70 S6 Kinase (Thr421/Ser424) Antibody detect endogenous levels of p70 S6 Kinase only when phosphorylated at the indicated sites. These antibodies also recognize p85 S6 Kinase when phosphorylated at the corresponding sites, Thr412 or Thr444/Ser447, respectively. Phospho-p70 S6 Kinase (Thr389) Antibody may also detect S6KII phosphorylated at Thr401.
Western Blotting
Western blot analysis of extracts from HeLa, NIH/3T3, PC12 and COS-7 cells using p70 S6 Kinase Antibody #9202.
Western Blotting
Western blot analysis of extracts from serum starved or serum-treated (20%) 293, NIH/3T3 and PC12 cells using Phospho-p70 S6 Kinase (Thr389) (108D2) Rabbit mAb #9234.
IP
Immunoprecipitation of extracts from serum-treated 293 cells under nondenaturing or denaturing conditions, using Phospho-p70 S6 Kinase (Thr421/Ser424) Antibody #9204. Western blot was then performed using the same antibody.
Source / Purification
Polyclonal antibodies are produced by immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr421/Ser424 of human p70 S6 kinase or a synthetic peptide derived from the sequence of human p70 S6 kinase (p70 S6 Kinase Antibody). Monoclonal antibodies are produced by immunizing immunizing rabbits with a synthetic phospho-peptide (KLH-coupled) corresponding to residues surrounding Thr389 of human p70 S6 Kinase. Antibodies are purified by Protein A and peptide affinity chromatography.
Background
p70 S6 kinase is a mitogen activated Ser/Thr protein kinase that is required for cell growth and G1 cell cycle progression (1,2). p70 S6 kinase phosphorylates the S6 protein of the 40S ribosomal subunit and is involved in translational control of 5' oligopyrimidine tract mRNAs (1). A second isoform, p85 S6 kinase, is derived from the same gene and is identical to p70 S6 kinase except for 23 extra residues at the amino terminus, which encode a nuclear localizing signal (1). Both isoforms lie on a mitogen activated signaling pathway downstream of phosphoinositide-3 kinase (PI-3K) and the target of rapamycin, FRAP/mTOR, a pathway distinct from the Ras/MAP kinase cascade (1). The activity of p70 S6 kinase is controlled by multiple phosphorylation events located within the catalytic, linker and pseudosubstrate domains (1). Phosphorylation of Thr229 in the catalytic domain and Thr389 in the linker domain are most critical for kinase function (1). Phosphorylation of Thr389, however, most closely correlates with p70 kinase activity in vivo (3). Prior phosphorylation of Thr389 is required for the action of phosphoinositide 3-dependent protein kinase 1 (PDK1) on Thr229 (4,5). Phosphorylation of this site is stimulated by growth factors such as insulin, EGF and FGF, as well as by serum and some G-protein-coupled receptor ligands, and is blocked by wortmannin, LY294002 (PI-3K inhibitor) and rapamycin (FRAP/mTOR inhibitor) (1,6,7). Ser411, Thr421 and Ser424 lie within a Ser-Pro-rich region located in the pseudosubstrate region (1). Phosphorylation at these sites is thought to activate p70 S6 kinase via relief of pseudosubstrate suppression (1,2). Another LY294002 and rapamycin sensitive phosphorylation site, Ser371, is an in vitro substrate for mTOR and correlates well with the activity of a partially rapamycin resistant mutant p70 S6 kinase (8).
- Pullen, N. and Thomas, G. (1997) FEBS Lett. 410, 78-82.
- Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
- Weng, Q.P. et al. (1998) J. Biol. Chem. 273, 16621-16629.
- Pullen, N. et al. (1998) Science 279, 707-710.
- Alessi, D.R. et al. (1998) Curr. Biol. 8, 69-81.
- Polakiewicz, R.D. et al. (1998) J. Biol. Chem. 273, 23534-23541.
- Fingar, D.C. et al. (2002) Genes Dev. 16, 1472-1487.
- Saitoh, M. et al. (2002) J. Biol. Chem. 277, 20104-20112.
Application References
- Aoki, M. et al. (2001) A role of the kinase mTOR in cellular transformation induced by the oncoproteins P3k and Akt. Proc. Natl. Acad. Sci. USA 98, 136-141. This article references the use of PhosphoPlus® p70 S6 Kinase (Thr389, Thr421/Ser424) Antibody Kit in the following applications: Western Blotting
- Kenerson, H. L. et al. (2002) Activated mammalian target of rapamycin pathway in the pathogenesis of tuberous sclerosis complex renal tumors. Cancer Res. 62, 5645-5650. This article references the use of PhosphoPlus® p70 S6 Kinase (Thr389, Thr421/Ser424) Antibody Kit in the following applications: IHC-P (paraffin)
- Sellers, L. A. et al. (2000) Receptor isoforms mediate opposing proliferative effects through gbetagamma-activated p38 or Akt pathways. Mol. Cell. Biol. 20, 5974-5985. This article references the use of PhosphoPlus® p70 S6 Kinase (Thr389, Thr421/Ser424) Antibody Kit in the following applications: Western Blotting
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
- 9205 Phospho-p70 S6 Kinase (Thr389) Antibody
- 9204 Phospho-p70 S6 Kinase (Thr421/Ser424) Antibody
- 9202 p70 S6 Kinase Antibody
- 9203 p70 S6 Kinase Control Cell Extracts
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 9208 Phospho-p70 S6 Kinase (Ser371) Antibody
- 9206 Phospho-p70 S6 Kinase (Thr389) (1A5) Mouse mAb
- 2217 S6 Ribosomal Protein (5G10) Rabbit mAb
- 2317 S6 Ribosomal Protein (54D2) Mouse mAb
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.