Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Stat6 (D3H4) Rabbit mAb #5397

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP ChIP H M R (Mk) (B) (Dg) (Pg) Endogenous 110 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  ChIP=Chromatin IP
Reactivity Key:  H=Human  M=Mouse  R=Rat  Mk=Monkey  B=Bovine  Dg=Dog  Pg=Pig
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Stat6 (D3H4) Rabbit mAb recognizes endogenous levels of total Stat6 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly582 of human Stat6 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Stat6 (D3H4) Rabbit mAb.

Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 Ramos cells starved overnight then treated with IL-4 (100 ng/ml, 30 min) and either 10 μl of Stat6 (D3H4) Rabbit mAb or 2 μl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using SimpleChIP® Human DMD Intron 2 Primers #7710, human MS4A1 promoter primers, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.

Background

Upon activation by Janus kinases, Stat6 translocates to the nucleus where it regulates cytokine-induced gene expression. Stat6 is activated via phosphorylation at Tyr641 and is required for responsiveness to IL-4 and IL-13 (1-4). In addition, Stat6 is activated by IFN-α in B cells, where it forms transcriptionally active complexes with Stat2 and p48 (5,6). Protein phosphatase 2A is also involved in regulation of IL-4-mediated Stat6 signaling (7).

  1. Nelms, K. et al. (1999) Ann. Rev. Immunol. 17, 701-738.
  2. Malabarba, M.G. et al. (1996) Biochem. J. 319, 865-872.
  3. Hou, J. et al. (1994) Science 265, 1701-1706.
  4. Quelle, F.W. et al. (1995) Mol. Cell. Biol. 15, 3336-3343.
  5. Takeda, K. et al. (1996) Nature 380, 627-630.
  6. Gupta, S. et al. (1999) J. Immunol. 163, 3834-3841.
  7. Woetmann, A. et al. (2003) J. Biol. Chem. 278, 2787-2791.

Application References

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Companion Products

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

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