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8209
PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet
Primary Antibodies
Antibody Duet

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet #8209

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PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 1

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 (upper), or p70 S6 Kinase (49D7) rabbit mAb #2708 (lower).

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 2

Immunohistochemical analysis of paraffin-embedded human colon carcinoma, using p70 S6 Kinase (49D7) Rabbit mAb.

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 3

Immunohistochemical analysis of paraffin-embedded human lung carcinoma, showing cytoplasmic and nuclear localization, using p70 S6 Kinase (49D7) Rabbit mAb.

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 4

Immunohistochemical analysis of paraffin-embedded human breast carcinoma, using p70 S6 Kinase (49D7) Rabbit mAb in the presence of control peptide (left) or p70 S6 Kinase Blocking Peptide #1205 (right).

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 5

Immunohistochemical analysis of paraffin-embedded mIMCD-3 Tsc1 (-/-) cell pellet (left, positive) or mIMCD-3 Tsc1/S6k1 (-/-) cell pellet (right, negative) using p70 S6 Kinase (49D7) Rabbit mAb. (Cell lines courtesy of Dr. Mario Pende, Inserm, Paris, France.)

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 6

Western blot analysis of extracts from PC12, NIH/3T3, and SK-N-MC cells, using p70 S6 Kinase (49D7) Rabbit mAb.

PhosphoPlus® p70 S6 Kinase (Thr389) Antibody Duet: Image 7

Western blot analysis of extracts from HeLa cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® p70/85 S6 Kinase siRNA I #6566 (+) or SignalSilence® p70/85 S6 Kinase siRNA II #6572 (+), using p70 S6 Kinase (49D7) Rabbit mAb #2708 and α-Tubulin (11H10) Rabbit mAb #2125. The p70 S6 Kinase (49D7) Rabbit mAb confirms silencing of p70/85 S6 kinase expression, while the α-Tubulin (11H10) Rabbit mAb is used to control for loading and specificity of p70/85 S6 kinase siRNA.

To Purchase # 8209S
Product # Size Price
8209S
1 Kit $ 514

Kit Usage Information

Protocols

Product Description

PhosphoPlus® Duets from Cell Signaling Technology (CST) provide a means to assess protein activation status. Each Duet contains an activation-state and total protein antibody to your target of interest. These antibodies have been selected from CST's product offering based upon superior performance in specified applications.

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).

  1. Pullen, N. and Thomas, G. (1997) FEBS Lett 410, 78-82.
  2. Dufner, A. and Thomas, G. (1999) Exp Cell Res 253, 100-9.
  3. Weng, Q.P. et al. (1998) J Biol Chem 273, 16621-9.
  4. Pullen, N. et al. (1998) Science 279, 707-10.
  5. Alessi, D.R. et al. (1998) Curr Biol 8, 69-81.
  6. Polakiewicz, R.D. et al. (1998) J Biol Chem 273, 23534-41.
  7. Fingar, D.C. et al. (2002) Genes Dev 16, 1472-87.
  8. Saitoh, M. et al. (2002) J Biol Chem 277, 20104-12.

Limited Uses

Except as otherwise expressly agreed in a writing signed by a legally authorized representative of CST, the following terms apply to Products provided by CST, its affiliates or its distributors. Any Customer's terms and conditions that are in addition to, or different from, those contained herein, unless separately accepted in writing by a legally authorized representative of CST, are rejected and are of no force or effect.

Products are labeled with For Research Use Only or a similar labeling statement and have not been approved, cleared, or licensed by the FDA or other regulatory foreign or domestic entity, for any purpose. Customer shall not use any Product for any diagnostic or therapeutic purpose, or otherwise in any manner that conflicts with its labeling statement. Products sold or licensed by CST are provided for Customer as the end-user and solely for research and development uses. Any use of Product for diagnostic, prophylactic or therapeutic purposes, or any purchase of Product for resale (alone or as a component) or other commercial purpose, requires a separate license from CST. Customer shall (a) not sell, license, loan, donate or otherwise transfer or make available any Product to any third party, whether alone or in combination with other materials, or use the Products to manufacture any commercial products, (b) not copy, modify, reverse engineer, decompile, disassemble or otherwise attempt to discover the underlying structure or technology of the Products, or use the Products for the purpose of developing any products or services that would compete with CST's products or services, (c) not alter or remove from the Products any trademarks, trade names, logos, patent or copyright notices or markings, (d) use the Products solely in accordance with CST's Product Terms of Sale and any applicable documentation, and (e) comply with any license, terms of service or similar agreement with respect to any third party products or services used by Customer in connection with the Products.

For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PhosphoPlus is a trademark of Cell Signaling Technology, Inc.