Revision 1
Cell Signaling Technology

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For Research Use Only. Not for Use in Diagnostic Procedures.
Product Includes Product # Quantity Mol. Wt Isotype/Source
Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (Alexa Fluor® 488 Conjugate) 9174 40 µl Rabbit IgG
Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) 4323 40 µl 86 kDa Rabbit IgG
Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) 3939 40 µl Rabbit IgG
Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) 2975 40 µl Rabbit IgG

Please visit cellsignal.com for individual component applications, species cross-reactivity, dilutions, protocols, and additional product information.

Description

The Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit provides an economical means to study the activation status of members of the Stat family of proteins without the need for a fluorescent secondary antibody.

Storage

Supplied in 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. Store at –20°C. Do not aliquot the antibody.

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 interferon response element (IRE) and gamma interferon-activated sequence (GAS) DNA elements, resulting in the transcriptional regulation of downstream genes (1,2). The remarkable range and specificity of responses regulated by the Stat family members is determined in part by the tissue-specific expression of different cytokine receptors, Jak and Stat family members (2,3), and by the combinatorial coupling of various Stat members to different receptors. Stat1 is activated in response to a large number of ligands (4) and is essential for responsiveness to IFN-α and IFN-γ (5,6). Stat3 is an important signaling molecule for many cytokines and growth factor receptors (4) and is required for murine fetal development (7). Stat5 is activated in response to a wide variety of ligands including IL-2, GM-CSF, growth hormone, and prolactin. Phosphorylation at Tyr694 is obligatory for Stat5 activation (8,9).

  1. Darnell, J.E. et al. (1994) Science 264, 1415-21.
  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, 13221-7.
  4. Heim, M.H. (1999) J Recept Signal Transduct Res 19, 75-120.
  5. Durbin, J.E. et al. (1996) Cell 84, 443-50.
  6. Meraz, M.A. et al. (1996) Cell 84, 431-42.
  7. Takeda, K. et al. (1997) Proc Natl Acad Sci U S A 94, 3801-4.
  8. Gouilleux, F. et al. (1994) EMBO J 13, 4361-9.
  9. Wakao, H. et al. (1994) EMBO J 13, 2182-91.

Background References

    Trademarks and Patents

    Cell Signaling Technology is a 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 1
    #12348

    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit

    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 1 Expand Image
    Confocal immunofluorescent analysis of Cos cells using GAPDH (14C10) Rabbit mAb (Alexa Fluor® 488 Conjugate) #3906 (green, left) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate (green, right). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).
    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 2 Expand Image
    Flow cytometric analysis of TF-1 cells untreated (blue) or treated with hGM-CSF (100 ng/ml, 15 min; green) using Phospho-Stat5 (Tyr694) (C71E5) Rabbit mAb (Alexa Fluor® 488 Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (dashed lines).
    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 3 Expand Image
    Flow cytometric analysis of Jurkat cells, untreated (blue) or treated with IFN-alpha #8927 (100ng/ml, 30min; green) using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) (solid lines) or a concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (dashed lines).
    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 4 Expand Image
    Flow cytometric analysis of Jurkat cells untreated (blue) or treated with IFN-alpha (100ng/ml, 5 min; green) using Phospho-Stat1 (Tyr701) (58D6) Rabbit mAb (Alexa Fluor® 488 Conjugate) (solid lines) or concentration-matched Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) #2975 (dashed lines).
    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 5 Expand Image
    Flow cytometric analysis of Jurkat cells, using Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) (red) compared to Phospho-S6 Ribosomal Protein (Ser235/236) (D57.2.2E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4803, untreated (blue) or IFN-α-treated (green).
    Stat Family Alexa Fluor® 488 Conjugated Antibody Sampler Kit: Image 6 Expand Image
    Flow cytometric analysis of Jurkat cells, using Rabbit (DA1E) mAb IgG XP® Isotype Control (Alexa Fluor® 488 Conjugate) (red) compared to Phospho-Akt (Ser473) (D9E) XP® Rabbit mAb (Alexa Fluor® 488 Conjugate) #4071, untreated (green) or LY294002- and wortmannin-treated (blue).