Product Pathways - PathScan ELISA
PathScan® Total p70 S6 Kinase Sandwich ELISA Antibody Pair #7039
|7039S||1 Kit (Reagents for 4 x 96 well plates)||---||In Stock||---|
|7039||carrier free and custom formulation / quantity||email request|
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|Kit Includes||Volume||Cap Color|
|p70 S6 Kinase Rabbit Capture Antibody (100X)||0.4 ml||Pink|
|Biotinylated p70 S6 Kinase Rabbit Detection Antibody (100X)||0.4 ml||Blue|
|HRP-linked Streptavidin (1000X)||0.04 ml||Clear|
Storage: Capture and detection antibodies are stored at 4°C. HRP-linked secondary reagent is stored at -20°C.
Cell Signaling Technology's PathScan® Total p70 S6 Kinase Sandwich ELISA Antibody Pair is being offered as an economical alternative to our PathScan® Total p70 S6 Kinase Sandwich ELISA Kit #7038. Capture and detection antibodies (100X stocks) and HRP-conjugated streptavidin (1000X stock) are supplied. Sufficient reagents are supplied for 4 x 96 well ELISAs. The p70 S6 kinase rabbit capture antibody is coated onto a 96 well microplate overnight in PBS. After blocking, cell lysates are added followed by a biotinylated p70 S6 kinase rabbit detection antibody and HRP-conjugated streptavidin. HRP substrate (TMB) is then added for color development. The magnitude of the absorbance for this developed color is proportional to the quantity of p70 S6 kinase.
Specificity / Sensitivity
For Antibody Pair specificity and sensitivity, please refer to the corresponding PathScan® Sandwich ELISA Kit. Note: This antibody pair detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
The relationship between the protein concentration of the lysate from untreated and IGF-1-treated MCF7 cells and the absorbance at 450 nm using the PathScan® Total p70 S6 Kinase Sandwich ELISA Antibody Pair #7039 is shown. MCF7 cells were treated with 100 ng/ml hIGF-I #8917 for 20 minutes at 37ºC and then lysed.
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).
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- Dufner, A. and Thomas, G. (1999) Exp. Cell Res. 253, 100-109.
- Weng, Q.P. et al. (1998) J. Biol. Chem. 273, 16621-16629.
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- 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.
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For Research Use Only. Not For Use In Diagnostic Procedures.
PathScan® is a trademark of Cell Signaling Technology, Inc.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.