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9133
Stat1/2/3/5 Control Cell Extracts
Experimental Controls
Cell Extract Kit

Stat1/2/3/5 Control Cell Extracts #9133

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  1. WB
Western Blotting Image 1: Stat1/2/3/5 Control Cell Extracts

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with IFN-alpha (100 ng/ml) for 5 min, using Stat1 (D1K9Y) Rabbit mAb #14994 (upper) and P-Stat1 (Tyr701) (58D6) Rabbit mAb #9167 (lower).

Western Blotting Image 2: Stat1/2/3/5 Control Cell Extracts

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with IFN-alpha (100 ng/ml) for 5 min, using Stat3 (D3Z2G) Rabbit mAb #12640 (upper) and P-Stat3 (Tyr705) (D3A7) XP Rabbit mAb #9145 (lower).

Western Blotting Image 3: Stat1/2/3/5 Control Cell Extracts

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with IFN-alpha (100 ng/ml) for 5 min, using Stat5 (D2O6Y) Rabbit mAb #94205 (upper) and P-Stat5 (Tyr694) (D47E7) Rabbit mAb #4322 (lower).

Product Usage Information

Boil for 3 minutes prior to use. Load 10 µl of phosphorylated and nonphosphorylated Stat1/2/3/5 Control Cell Extracts per lane.

Storage

Supplied in SDS Sample Buffer: 62.5 mM Tris-HCl (pH 6.8 at 25°C), 2% w/v SDS, 10% glycerol, 50 mM DTT, 0.01% w/v bromophenol blue or phenol red. Store at –20°C. For long term, store at –80°C.

Product Description

Stat1/2/3/5 Control Cell Extracts (HeLa untreated): Total cell extracts from serum-starved HeLa cells serve as a negative control. Supplied in SDS Sample Buffer.

Stat1/2/3/5 Control Cell Extracts (HeLa + IFN-alpha): Total cell extracts from serum-starved HeLa cells treated with 100 ng/ml interferon-alpha for 5 minutes serve as a positive control. Supplied in SDS Sample Buffer.

Background

Jaks (Janus Kinases) and Stats (Signal Transducers and Activators of Transcription) are utilized by receptors for a wide variety of ligands including cytokines, hormones, growth factors and neurotransmitters. Jaks, activated via autophosphorylation following ligand-induced receptor aggregation, phosphorylate tyrosine residues on associated receptors, Stat molecules and other downstream signaling proteins (1,2). The phosphorylation of Stat proteins at conserved tyrosine residues activates SH2-mediated dimerization followed rapidly by nuclear translocation. Stat dimers bind to IRE (interferon response element) and GAS (gamma interferon-activated sequence) DNA elements, resulting in the transcriptional regulation of downstream genes (1,2). The remarkable range and specificity of responses regulated by the Stats is determined in part by the tissue-specific expression of different cytokine receptors, Jaks and Stats (2,3), and by the combinatorial coupling of various Stat members to different receptors. Serine phosphorylation in the carboxy-terminal transcriptional activation domain has been shown to regulate the function of Stat1, -2, -3, -4 and -5 (1). Phosphorylation of Stat3 at Ser727 via MAPK or mTOR pathways is required for optimal transcriptional activation in response to growth factors and cytokines including IFN-gamma and CNTF (4,5). Jak/Stat pathways also play important roles in oncogenesis, tumor progression, angiogenesis, cell motility, immune responses and stem cell differentiation (6-11).

  1. Darnell Jr., J. et al. (1994) Science 264, 1415-1421.
  2. Leonard, W.J. and O'Shea, J.J. (1998) Annu Rev Immunol 16, 293-322.
  3. Caldenhoven, E. et al. (1996) J. Biol. Chem. 271, 13221-13227.
  4. Wen, Z. et al. (1995) Cell 82, 241-250.
  5. Yokogami, K. et al. (2000) Curr. Biol. 10, 47-50.
  6. Lim, C.P. and Cao, X. (1999) J. Biol. Chem. 274, 31055-31061.
  7. Bromberg, J. F. et al. (1999) Cell 98, 295-303.
  8. Su, L. et al. (1999) J. Biol. Chem. 274, 31770-31774.
  9. Dentelli, P. et al. (1999) J. Immunol. 163, 2151-2159.
  10. Cattaneo, E. et al. (1999) Trends Neurosci. 22, 365-369.
  11. Frank, D.A. (1999) Mol. Med. 5, 432-456.

Limited Uses

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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.
To Purchase # 9133S
Product # Size Price
9133S
100 µl  (10 western blots) N/A

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