Cell Signaling Technology

Product Pathways - Development

Cleaved Notch1 (Val1744) Antibody #2421

Applications Reactivity Sensitivity MW (kDa) Source
W IP H M R Mk Endogenous 110 Rabbit

Applications Key:  W=Western Blotting  IP=Immunoprecipitation
Reactivity Key:  H=Human  M=Mouse  R=Rat  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

Cleaved Notch1 (Val1744) Antibody detects endogenous levels of the Notch1 intracellular domain (NICD) only when released by cleavage between Gly1753 and Val1754 (equivalent to Gly1743/Val1744 of murine Notch1). This antibody does not recognize full-length Notch1 or Notch1 cleaved at other positions. This antibody may cross-react with overexpressed cleaved Notch2, 3 and 4.

Source / Purification

Polyclonal antibodies are produced by immunizing animals with a synthetic peptide corresponding to the sequence at the Val1754 cleavage site in human Notch 1 (equivalent to Val1744 in mouse Notch1). Antibodies are purified by protein A and peptide affinity chromatography.

Western Blotting

Western Blotting

Western blot analysis of extracts from 293 cells transfected with full-length Notch1, either untreated or treated with 5 mM EDTA for 15 minutes, and MOLT-4 extracts, using Cleaved Notch 1 (Val1744) Antibody.

Background

Notch proteins (Notch1-4) are a family of transmembrane receptors that play important roles in development and the determination of cell fate (1). Mature Notch receptors are processed and assembled as heterodimeric proteins, with each dimer comprised of a large extracellular ligand-binding domain, a single-pass transmembrane domain, and a smaller cytoplasmic subunit (Notch intracellular domain, NICD) (2). Binding of Notch receptors to ligands of the Delta-Serrate-Lag2 (DSL) family triggers heterodimer dissociation, exposing the receptors to proteolytic cleavages; these result in release of the NICD, which translocates to the nucleus and activates transcription of downstream target genes (3-4).

The NICD of murine Notch1 is released (activated) by cleavage between Gly1743 and Val1744 (corresponding to Gly1753/Val1754 in human Notch1) (3, 4). Mutations that result in constitutive activation of Notch1 are associated with many different cancers, including a majority of cases of T cell acute lymphoblastic leukemia (T-ALL). Activation may be due to mutations in Notch1 itself, or in components of the ubiquitin ligase complex that negatively regulates the Notch signaling pathway (5-6).

  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. Weng, A.P. et al. (2004) Science 306, 269-71.
  6. Thompson, B.J. et al. (2007) J Exp Med 204, 1825-35.

Application References

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

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