Cell Signaling Technology

Product Pathways - Jak/Stat Pathway

CISH (D4D9) Rabbit mAb #8731

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IF-IC F H M (Mk) Endogenous 32, 37 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse  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

CISH (D4D9) Rabbit mAb recognizes endogenous levels of total CISH protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Pro176 of human CISH protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from NK-92 cells, untreated (-) or treated (+) with Human Interleukin-2 (hIL-2) #8907 (10 ng/ml, overnight), or BaF3 cells, untreated (-) or treated (+) with Mouse Interleukin-3 (mIL-3) #8923 (10 ng/ml, 6 hr), using CISH (D4D9) Rabbit mAb (upper) and β-Actin (D6A8) Rabbit mAb #8457 (lower).

Western Blotting

Western Blotting

Western blot analysis of extracts from 293T cells, mock transfected (-) or transfected with a construct expressing full-length human CISH (hCISH, +), using CISH (D4D9) Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of NK-92 cells, IL-2 starved for 5 hrs (blue) or IL-2 starved for 5 hrs followed by additon of hIL-2 #8907 overnight (green), using CISH (D4D9) Rabbit mAb (green).


IF-IC

IF-IC

Confocal immunofluorescent analysis of NK-92 cells, IL-2 starved for 5 hrs (left) or IL-2 starved for 5 hrs followed by addition of hIL-2 #8907 overnight (right), using CISH (D4D9) Rabbit mAb (green). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

The suppressor of cytokine signaling (SOCS) family members are negative regulators of cytokine signal transduction that inhibit the Jak/Stat pathway (1-3). The SOCS family consists of at least 8 members including the originally identified cytokine-inducible SH2-containing protein (CIS1), as well as SOCS1-7. Each SOCS family member contains a central SH2 domain and a conserved carboxy-terminal motif designated as the SOCS box. These proteins are important regulators of cytokine signaling, proliferation, differentiation, and immune responses.

CISH/CIS1, the first described member of the SOCS family, is induced by a number of cytokines including IL-2, IL-3, GM-CSF, and EPO (4). The CISH protein appears as a doublet around 32 and 37 kDa, the nature of which is unknown (4). CISH binds to phosphorylated cytokine receptors and can inhibit Stat5 activity (4-6). Expression of CISH is regulated by Stat5, thereby providing feedback modulation (5). Transgenic mice overexpressing CISH display phenotypes similar to Stat5 knockouts, including defects in mammary gland development and in T and NK cell regulation (6). Research studies have shown that polymorphisms within the CISH gene are associated with susceptibility to infectious diseases (7).

  1. Alexander, W.S. et al. (1999) J Leukoc Biol 66, 588-92.
  2. Chen, X.P. et al. (2000) Immunity 13, 287-90.
  3. Hilton, D.J. et al. (1998) Proc Natl Acad Sci USA 95, 114-9.
  4. Yoshimura, A. et al. (1995) EMBO J 14, 2816-26.
  5. Matsumoto, A. et al. (1997) Blood 89, 3148-54.
  6. Matsumoto, A. et al. (1999) Mol Cell Biol 19, 6396-407.
  7. Khor, C.C. et al. (2010) N Engl J Med 362, 2092-101.

Application References

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Companion Products

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

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