Cell Signaling Technology

Product Pathways - Wnt / Hedgehog / Notch

Notch3 (8G5) Rat mAb #3446

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H R Endogenous 90, 270 Rat IgG2a

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  R=Rat
Species cross-reactivity is determined by western blot. Species enclosed in parentheses are predicted to react based on 100% sequence homology.

Protocols

Specificity / Sensitivity

Notch3 (8G5) Rat mAb detects endogenous levels of total notch3 protein. It recognizes the full-length (270 kDa) and the extracellular truncated fragment containing a short extracellular region, the transmembrane domain and the intracellular region (90 kDa).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a fusion protein corresponding to intracellular residues of notch3.

Western Blotting

Western Blotting

Western blot analysis of extracts from HBP-ALL and A204 cells using Notch3 (8G5) Rat mAb. Full-length (FL) and cleaved notch3 proteins are indicated.

Background

Notch1 is a transmembrane protein functioning in development and the determination of cell fate (1). During maturation, the notch molecule is cleaved by a furin-like convertase at its extracellular domain (2). Upon binding to a ligand such as Delta1, or upon extracellular calcium depletion, the carboxy-terminal notch1 fragment is released and further cleaved between Gly1743 and Val1744 (3,4). The resulting activated cytosolic fragment translocates to the nucleus where it activates transcription.

Notch3 is a member of notch family that is processed in a similar way to notch1 (5). It is expressed primarily in arterial smooth muscle cells (SMC). Mutations altering the number of cysteine residues in the notch3 extracellular region are associated with cerebral autosomal dominant arteriopathy with subcortical infarcts and leukoencephalopathy (CADASIL), a hereditary angiopathy leading to strokes and dementia in adults (6-8). Recent studies indicate that notch3 is overexpressed in many types of cancers (9-11).

  1. Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6.
  2. Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6.
  3. Schroeter, E.H. et al. (1998) Nature 393, 382-6.
  4. Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35.
  5. Baron, M. (2003) Semin Cell Dev Biol 14, 113-9.
  6. Kalimo, H. et al. (2002) Brain Pathol 12, 371-84.
  7. Karlström, H. et al. (2002) Proc Natl Acad Sci USA 99, 17119-24.
  8. Monet, M. et al. (2007) Hum Mol Genet 16, 982-92.
  9. Park, J.T. et al. (2006) Cancer Res 66, 6312-8.
  10. Gramantieri, L. et al. (2007) Liver Int 27, 997-1007.
  11. Yamaguchi, N. et al. (2008) Cancer Res 68, 1881-8.

Application References

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Companion Products

This product is intended for research purposes only. The product is not intended to be used for therapeutic or diagnostic purposes in humans or animals.

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