Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb #8826

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P ChIP H M R (Mk) (B) Endogenous 91 Rabbit IgG

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

Protocols

Specificity / Sensitivity

Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb recognizes endogenous levels of Stat1 protein only when phosphorylated at Ser727.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Ser727 of human Stat1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines, untreated (-) or UV-treated (2 hr recovery; +), using Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from UV-treated HeLa cells, untreated (-) or CIP and λ phosphatase-treated (+), using Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb (upper) or Stat1 (42H3) Rabbit mAb #9175 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from HeLa cells, untreated (-) or treated with Human Interferon-γ (hIFN-γ) #8901 (100 ng/ml, 30 min; +), using Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb (upper) or Stat1 (42H3) Rabbit mAb #9175 (lower).


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human lymphoma, control (left) or λ phosphatase-treated (right) using Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human ovarian carcinoma using Phospho-Stat1 (Ser727) (D3B7) Rabbit mAb.


Chromatin IP

Chromatin IP

Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 HT-1080 cells treated with IFN-γ (50 ng/ml) for 30 minutes and either 10 μl of Phospho-Stat1 (Ser727) (D3B7) 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 human IRF-1 promoter primers, SimpleChIP® Human TAP1 Promoter Primers #5148, 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

The Stat1 transcription factor is activated in response to a large number of ligands (1) and is essential for responsiveness to IFN-α and IFN-γ (2,3). Phosphorylation of Stat1 at Tyr701 induces Stat1 dimerization, nuclear translocation, and DNA binding (4). Stat1 protein exists as a pair of isoforms, Stat1α (91 kDa) and the splice variant Stat1β (84 kDa). In most cells, both isoforms are activated by IFN-α, but only Stat1α is activated by IFN-γ. The inappropriate activation of Stat1 occurs in many tumors (5). In addition to tyrosine phosphorylation, Stat1 is also phosphorylated at Ser727 through a p38 mitogen-activated protein kinase (MAPK)-dependent pathway in response to IFN-α and other cellular stresses (6). Serine phosphorylation may be required for the maximal induction of Stat1-mediated gene activation.

  1. Heim, M.H. (1999) J. Recept. Signal. Transduct. Res. 19, 75-120.
  2. Durbin, J.E. et al. (1996) Cell 84, 443-450.
  3. Meraz, M.A. et al. (1996) Cell 84, 431-442.
  4. Ihle, J.N. et al. (1994) Trends Biochem. Sci. 19, 222-227.
  5. Frank, D.A. (1999) Mol. Med. 5, 432-456.
  6. Wen, Z. et al. (1995) Cell 82, 241-250.

Application References

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

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