Cell Signaling Technology

Product Pathways - Wnt / Hedgehog / Notch

Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb #4147

Applications Reactivity Sensitivity MW (kDa) Isotype
W IP H M R Endogenous 110 Rabbit IgG

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

Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb detects endogenous levels of the cytosolic domain of Notch1 only when cleaved between Gly1743 and Val1744. The antibody does not recognize full length Notch1 or Notch1 cleaved at other positions. It detects Cleaved Notch1 with different sizes in different cell lines due to mutations at the distant C-termini of Notch1 (see refs 6 and 7).

Source / Purification

Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence at the Val1744 cleavage site of human Notch1.

Western Blotting

Western Blotting

Western blot analysis of extracts from various cell lines using Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb (upper) or Notch1 (D1E11) XP® Rabbit mAb #3608 (lower).

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.

Constitutively activated Notch1 signaling is associated with the majority of cases of T cell acute lymphoblastic leukemia (T-ALL). The activation is either due to mutations in Notch1 itself or in the components of ubiquitin ligase complex, namely FBW7 (5-7).

  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.
  7. Thompson, B.J. et al. (2007) J Exp Med 204, 1825-35.

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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