Product Pathways - Jak/Stat Pathway

Phospho-Stat Antibody Sampler Kit #9914

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Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IHC-F=Immunohistochemistry (Frozen)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine

Specificity / Sensitivity

Each phospho-Stat antibody in the kit recognizes only the phosphorylated form of its specific target. Antibodies 9131 and 9351 do cross-react with EGFR.

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 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 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) Rabbit mAb #9145. Note that the basal phospho-Stat3 in A431 is detected by the antibody.

Western Blotting

Western blot analysis of extracts from IFN-alpha treated (100 ng/ml) and untreated HeLa cells, using Phospho-Stat1 (Tyr701) Antibody #9171 .

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

IC-ABC

Phospho-Stat1 (Tyr701) Antibody #9171 allows in situ detection and sub-cellular resolution of Stat1 phosphorylation and nuclear translocation induced by IFN-alpha.

Table

Table

Source / Purification

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

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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Companion Products

• 7074 Anti-rabbit IgG, HRP-linked Antibody
• 7003 20X LumiGLO^® Reagent and 20X Peroxide
• 9132 Stat3 Antibody
• 9133 Stat3 Control Cell Extracts
• 9136 Phospho-Stat3 (Ser727) (6E4) Mouse mAb
• 9939 Stat Antibody Sampler Kit
• 9175 Stat1 (42H3) Rabbit mAb (Human Specific)
• 9363 Stat5 Antibody
• 9173 Stat1 Control Cell Extracts
• 9353 Stat5 Control Cell Extracts
• 4594 Stat2 Antibody
• 9362 Stat6 Antibody
• 9177 Phospho-Stat1 (Ser727) Antibody