Cell Signaling Technology
XP Monoclonal Antibody

Product Pathways - Lymphocyte Signaling

NFAT1 (D43B1) XP® Rabbit mAb #5861

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP IHC-P IF-IC F H M Endogenous 140 Rabbit IgG

Applications Key:  W=Western Blotting  IP=Immunoprecipitation  IHC-P=Immunohistochemistry (Paraffin)  IF-IC=Immunofluorescence (Immunocytochemistry)  F=Flow Cytometry
Reactivity Key:  H=Human  M=Mouse
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

* Product-specific protocol.

Specificity / Sensitivity

NFAT1 (D43B1) XP® Rabbit mAb detects endogenous levels of total NFAT1 protein.

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to residues surrounding Gly87 of human NFAT1 protein.

Western Blotting

Western Blotting

Western blot analysis of extracts from Ramos, Raji, and EL4 cells using NFAT1 (D43B1) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human breast carcinoma using NFAT1 (D43B1) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human colon carcinoma using NFAT1 (D43B1) XP® Rabbit mAb.


IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded human T-cell lymphoma using NFAT1 (D43B1) XP® Rabbit mAb.

IHC-P (paraffin)

IHC-P (paraffin)

Immunohistochemical analysis of paraffin-embedded cell pellets, Jurkat (left) or LNCaP (right), using NFAT1 (D43B1) XP® Rabbit mAb.

Flow Cytometry

Flow Cytometry

Flow cytometric analysis of HCT 116 using NFAT1 (D43B1) XP® Rabbit mAb (blue) compared to concentration matched Rabbit (DA1E) mAb IgG XP® Isotype Control #3900 (red).


IF-IC

IF-IC

Confocal immunofluorescent analysis of MCF7 cells, untreated (left) or treated with Ionomycin #9995 (1 μM, 1 hr; right), using NFAT1 (D43B1) XP® Rabbit mAb (green). Actin filaments were labeled with DY-554 phalloidin (red). Blue pseudocolor = DRAQ5® #4084 (fluorescent DNA dye).

Background

The NFAT (nuclear factor of activated T cells) family of proteins consists of NFAT1 (NFATc2 or NFATp), NFAT2 (NFATc1 or NFATc), NFAT3 (NFATc4), and NFAT4 (NFATc3 or NFATx). All members of this family are transcription factors with a Rel homology domain and regulate gene transcription in concert with AP-1 (Jun/Fos) to orchestrate an effective immune response (1,2). NFAT proteins are predominantly expressed in cells of the immune system, but are also expressed in skeletal muscle, keratinocytes, and adipocytes, regulating cell differentiation programs in these cells (3). In resting cells, NFAT proteins are heavily phosphorylated and localized in the cytoplasm. Increased intracellular calcium concentrations activate the calcium/calmodulin-dependent serine phosphatase calcineurin, which dephosphorylates NFAT proteins, resulting in their subsequent translocation to the nucleus (2). Termination of NFAT signaling occurs upon declining calcium concentrations and phosphorylation of NFAT by kinases such as GSK-3 or CK1 (3,4). Cyclosporin A and FK506 are immunosuppressive drugs that inhibit calcineurin and thus retain NFAT proteins in the cytoplasm (5).

  1. Northrop, J.P. et al. (1993) J. Biol. Chem. 268, 2917-2923.
  2. Hogan, P.G. et al. (2003) Genes Dev. 17, 2205-2232.
  3. Crabtree, G.R. and Olson, E.N. (2002) Cell 109, S67-S79.
  4. Okamura, H. et al. (2004) Mol. Cell. Biol. 24, 4184-4195.
  5. Shaw, K.T. et al. (1995) Proc. Natl. Acad. Sci. USA 92, 11205-11209.

Application References

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Companion Products

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

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