Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Jak/Stat Pathway

PIAS3 (D5F9) XP® Rabbit mAb #9042

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC H (Mk) Endogenous 65-75 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)
Reactivity Key:  H=Human  Mk=Monkey
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

PIAS3 (D5F9) XP® Rabbit mAb recognizes endogenous levels of total PIAS3 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro590 of human PIAS3 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from RD cells, transfected with 100 nM SignalSilence® Control siRNA (Unconjugated) #6568 (-), SignalSilence® PIAS3 siRNA I #9073 (+), or SignalSilence® PIAS3 siRNA II #9031 (+), using PIAS3 (D5F9) XP® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). The PIAS3 (D5F9) XP® Rabbit mAb confirms silencing of PIAS3 expression, while the β-Actin (D6A8) Rabbit mAb is used as a loading control.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using PIAS3 (D5F9) XP® Rabbit mAb.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human PIAS3 (hPIAS3; +), using PIAS3 (D5F9) XP® Rabbit mAb.


IF-IC

IF-IC

Confocal immunofluorescent analysis of A-204 (positive, left) and Hep G2 (low expression, right) cells using PIAS3 (D5F9) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red).

Background

The protein inhibitor of activated Stat (PIAS) proteins, which include PIAS1, PIAS3, PIASx, and PIASy, were originally characterized based on their interaction with the Stat family of transcription factors (1,2). PIAS1, PIAS3, and PIASx interact with and repress Stat1, Stat3, and Stat4, respectively (1-3). Deletion of PIAS1 leads to inhibition of interferon-inducible genes and increased protection against infection (4). The PIAS family contains a conserved RING domain that has been linked to a function as a small ubiquitin-related modifier (SUMO) ligase, coupling the SUMO conjugating enzyme Ubc9 with its substrate proteins (5,6). Numerous studies have now shown that PIAS family members can regulate the activity of transcription factors through distinct mechanisms, including NF-κB (7,8), c-Jun, p53 (5,9), Oct-4 (10), and Smads (11,12). The activity of PIAS1 is regulated by both phosphorylation and arginine methylation. Inflammatory stimuli can induce IKK-mediated phosphorylation of PIAS1 at Ser90, which is required for its activity (13). In addition, PRMT1 induces arginine methylation of PIAS1 at Arg303 following interferon treatment and is associated with its repressive activity on Stat1 (14).

  1. Liu, B. et al. (1998) Proc Natl Acad Sci USA 95, 10626-31.
  2. Chung, C.D. et al. (1997) Science 278, 1803-5.
  3. Arora, T. et al. (2003) J Biol Chem 278, 21327-30.
  4. Liu, B. et al. (2004) Nat Immunol 5, 891-8.
  5. Schmidt, D. and Müller, S. (2002) Proc Natl Acad Sci USA 99, 2872-7.
  6. Kotaja, N. et al. (2002) Mol Cell Biol 22, 5222-34.
  7. Liu, B. et al. (2005) Mol Cell Biol 25, 1113-23.
  8. Tahk, S. et al. (2007) Proc Natl Acad Sci USA 104, 11643-8.
  9. Bischof, O. et al. (2006) Mol Cell 22, 783-94.
  10. Tolkunova, E. et al. (2007) J Mol Biol 374, 1200-12.
  11. Long, J. et al. (2004) Proc Natl Acad Sci USA 101, 99-104.
  12. Murdoch, R.N. and Edwards, T. (1992) Biochem Int 28, 1029-37.
  13. Liu, B. et al. (2007) Cell 129, 903-14.
  14. Weber, S. et al. (2009) Genes Dev 23, 118-32.

Application References

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Companion Products

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

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