Revision 5
#7149
Store at +4C
PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit
1 Kit
(96 assays)
Species Cross Reactivity:
H M
UniProt ID:
#P40763
Entrez-Gene Id:
#6774
877-616-CELL (2355)
877-678-TECH (8324)
3 Trask Lane | Danvers | Massachusetts | 01923 | USA
For Research Use Only. Not for Use in Diagnostic Procedures.
| Product Includes | Product # | Quantity | Color | Storage Temp |
|---|---|---|---|---|
| Anti-mouse IgG, HRP-linked Antibody (ELISA Formulated) | 13304 | 1 ea | Red (Lyophilized) | UNKNOWN |
| Phospho-Stat3 (Tyr705) Mouse Detection mAb | 13149 | 1 ea | Green (Lyophilized) | UNKNOWN |
| Stat3 Rabbit mAb Coated Microwells | 52093 | 96 tests | +4C | |
| Detection Antibody Diluent | 13339 | 5.5 ml | Green | +4C |
| HRP Diluent | 13515 | 5.5 ml | Red | +4C |
| Luminol/Enhancer Solution | 84850 | 3 ml | RT | |
| Stable Peroxide Buffer | 42552 | 3 ml | RT | |
| Sealing Tape | 54503 | 2 ea | +4C | |
| ELISA Wash Buffer (20X) | 9801 | 25 ml | +4C | |
| ELISA Sample Diluent | 11083 | 25 ml | Blue | +4C |
| Cell Lysis Buffer (10X) | 9803 | 15 ml | -20C |
*The microwell plate is supplied as 12 8-well modules - Each module is designed to break apart for 8 tests.
Description
The PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit is a solid phase sandwich enzyme-linked immunosorbent assay (ELISA) that detects endogenous levels of phospho-Stat3 (Tyr705) protein with a chemiluminescent readout. Chemiluminescent ELISAs often have a wider dynamic range and higher sensitivity than conventional chromogenic detection. This chemiluminescent ELISA, which is offered in low volume microplates, shows increased signal and sensitivity while using smaller sample volume. A Stat3 antibody has been coated on the microwells. After incubation with cell lysates, phospho and nonphospho-Stat3 proteins are captured by the coated antibody. Following extensive washing, a phospho-Stat3 mouse mAb is added to detect the captured phospho-Stat3 (Tyr705) protein. Anti-mouse IgG, HRP-linked antibody is then used to recognize the bound detection antibody. Chemiluminescent reagent is added for signal development. The magnitude of light emission, measured in relative light units (RLU), is proportional to the quantity of phospho-Stat3 (Tyr705) protein.
*Antibodies in this kit are custom formulations specific to kit.
Specificity/Sensitivity
PathScan® Phospho-Stat3 (Tyr705) Chemiluminescent Sandwich ELISA Kit detects endogenous levels of phospho-Stat3 (Tyr705) in human and mouse cells. This kit detects proteins from the indicated species, as determined through in-house testing, but may also detect homologous proteins from other species.
Background
The Stat3 transcription factor is an important signaling molecule for many cytokines and growth factor receptors (1) and is required for murine fetal development (2). Research studies have shown that Stat3 is constitutively activated in a number of human tumors (3,4) and possesses oncogenic potential (5) and anti-apoptotic activities (3). Stat3 is activated by phosphorylation at Tyr705, which induces dimerization, nuclear translocation, and DNA binding (6,7). Transcriptional activation seems to be regulated by phosphorylation at Ser727 through the MAPK or mTOR pathways (8,9). Stat3 isoform expression appears to reflect biological function as the relative expression levels of Stat3α (86 kDa) and Stat3β (79 kDa) depend on cell type, ligand exposure, or cell maturation stage (10). It is notable that Stat3β lacks the serine phosphorylation site within the carboxy-terminal transcriptional activation domain (8).
Background References
- Heim, M.H. (2001) J Recept Signal Transduct Res 19, 75-120.
- Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
- Catlett-Falcone, R. et al. (1999) Immunity 10, 105-15.
- Garcia, R. and Jove, R. (1998) J Biomed Sci 5, 79-85.
- Bromberg, J.F. et al. (1999) Cell 98, 295-303.
- Darnell, J.E. et al. (1994) Science 264, 1415-21.
- Ihle, J.N. (1995) Nature 377, 591-4.
- Wen, Z. et al. (1995) Cell 82, 241-50.
- Yokogami, K. et al. (2000) Curr Biol 10, 47-50.
- Biethahn, S. et al. (1999) Exp Hematol 27, 885-94.
Cross-Reactivity Key
H: Human M: Mouse R: Rat Hm: Hamster Mk: Monkey Vir: Virus Mi: Mink C: Chicken Dm: D. melanogaster X: Xenopus Z: Zebrafish B: Bovine Dg: Dog Pg: Pig Sc: S. cerevisiae Ce: C. elegans Hr: Horse GP: Guinea Pig Rab: Rabbit G: Goat All: All Species Expected
Trademarks and Patents
Cell Signaling Technology is a trademark of Cell Signaling Technology, Inc.
PathScan is a registered trademark of Cell Signaling Technology, Inc.
All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.
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 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 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.
Revision 5
Relationship between protein concentration of lysates from untreated and IFN-α treated HeLa cells and immediate light generation with chemiluminescent substrate is shown. Cells (75% confluence) were treated with IFN-α (100 ng/ml) and lysed after incubation at 37ºC for 10 minutes. Graph inset corresponding to the shaded area shows high sensitivity and a linear response at the low protein concentration range.
