Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat Antibody Sampler Kit #9914

Kit Includes Quantity Applications Reactivity MW (kDa) Isotype
Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb #7649 40 µl W IP IF-IC F ChIP H M R (Mk) 84, 91 Rabbit IgG
Phospho-Stat2 (Tyr690) Antibody #4441 40 µl W H (R) (B) 113 Rabbit
Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145 40 µl W IP IHC-P IHC-F IF-IC F ChIP H M R Mk (Hm) (B) (Pg) (Hr) 79, 86 Rabbit IgG
Phospho-Stat3 (Ser727) Antibody #9134 40 µl W IP IF-IC ChIP H M R (Hm) (Mk) (B) (Dg) (Pg) (Hr) 86 Rabbit
Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb #9359 40 µl W IP IHC-P F H M (R) (Mk) (B) 90 Rabbit IgG
Phospho-Stat6 (Tyr641) Antibody #9361 40 µl W IP IF-IC F H (B) 110 Rabbit
Anti-rabbit IgG, HRP-linked Antibody #7074 100 µl Goat

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Hm=Hamster  Mk=Monkey  B=Bovine  Dg=Dog  Pg=Pig  Hr=Horse
Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Specificity / Sensitivity

Each phospho-Stat antibody in the kit recognizes only the phosphorylated form of its specific target.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated or IFN-a-treated (100 ng/ml,15 minutes), using Phospho-Stat2 (Tyr690) Antibody or total stat2 antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa, A20, and PC-12 cells, untreated or treated with Human Interferon-α1 (hIFN-α1) #8927 (10 ng/ml, 30 min), using Phospho-Stat1 (Tyr701) (D4A7) Rabbit mAb #7649 (upper) or Stat1 Antibody #9172 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from EGF treated (100 ng/ml) A431 cells, using Phospho-Stat3 (Ser727) Antibody #9134 and Stat3 Antibody #9132.


Western Blotting

Western Blotting

Western blot analysis of extracts from IFN-a treated Jurkat cells and HeLa cells (left), as well as EGF treated A431 cells (right), using Phospho-Stat3 (Tyr705) (D3A7) XP® Rabbit mAb #9145. Note that the basal phospho-Stat3 in A431 is detected by the antibody.

Western Blotting

Western Blotting

Western blot analysis of extracts from K562 cells, untreated or treated with Gleevec, using Phospho-Stat5 (Tyr694) (C11C5) Rabbit mAb #9359 (top) and total Stat5 (3H7) Rabbit mAb #9358 (bottom).

Western Blotting

Western Blotting

Western blot analysis of extracts from IL-4 treated (10 ng/ml) Daudi cells, using Phospho-Stat6 (Tyr641) Antibody #9361 or control Stat6 antibody.


Description

The Phospho-Stat Pathway Sampler Kit provides an economical means to evaluate the activation status of Stat molecules, including the phosphorylation of Stat1 at Tyr701, Stat2 at Tyr690, Stat3 at Tyr705/Ser727, Stat5 at Tyr694 and Stat6 at Tyr641. The kit includes enough primary and secondary antibody to perform four Western blot experiments.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic phosphopeptides corresponding to residues surrounding Tyr690 of human Stat2, Ser727 of mouse Stat3, Tyr694 of mouse Stat5 or Tyr641 of human Stat6. Antibodies are purified by protein A and peptide affinity chromatography. Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Tyr701 of human Stat1 protein or Tyr705 of mouse Stat3.

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.

Application References

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Protocols

* Product-specific protocol.

Companion Products

Selected rabbit monoclonal antibodies are produced under license (granting certain rights including those under U. S. Patent No. 5,675,063 and/or U.S.S.N. 11/476,277) from Epitomics, Inc.

For Research Use Only. Not For Use In Diagnostic Procedures.

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