Product Pathways - Development
Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb #4147
|4147S||100 µl (10 western blots)||---||In Stock||---|
|4147P||40 µl (4 western blots)||---||In Stock||---|
|4147||carrier free and custom formulation / quantity||email request|
Already purchased this product? Write a Review.
|W||1:1000||Human, Mouse, Rat||Endogenous||110||Rabbit IgG|
Species cross-reactivity is determined by western blot.
Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP
Specificity / Sensitivity
Cleaved Notch1 (V1744) (D3B8) Rabbit mAb 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). The antibody does not recognize full-length Notch1 or Notch1 cleaved at other positions. The size of the NICD varies among cell lines due to mutations in the Notch1 C-terminus (6).
Source / Purification
Monoclonal antibody is produced by immunizing animals with a synthetic peptide corresponding to the sequence at the Val1754 cleavage site in human Notch1 (equivalent to Val1744 in mouse Notch1).
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).
CUTLL1 cells were cultured in media with γ-secretase inhibitor (1μM) for 3 days and then either harvested immediately (left panel) or washed and cultured in fresh media for 3h (right panel). Chromatin immunoprecipitations were performed with cross-linked chromatin from 4 x 106 cells and 2.5 µl of Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb or 2 µl of Normal Rabbit IgG #2729 using SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads) #9003. The enriched DNA was quantified by real-time PCR using human HES1 promoter primers, SimpleChIP® Human HES4 Promoter Primers #7273, and SimpleChIP® Human α Satellite Repeat Primers #4486. The amount of immunoprecipitated DNA in each sample is represented as signal relative to the total amount of input chromatin, which is equivalent to one.
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).
- Artavanis-Tsakonas, S. et al. (1999) Science 284, 770-6.
- Chan, Y.M. and Jan, Y.N. (1998) Cell 94, 423-6.
- Schroeter, E.H. et al. (1998) Nature 393, 382-6.
- Rand, M.D. et al. (2000) Mol Cell Biol 20, 1825-35.
- Weng, A.P. et al. (2004) Science 306, 269-71.
- Thompson, B.J. et al. (2007) J Exp Med 204, 1825-35.
- Treanor, L.M. et al. (2011) Blood 117, 5453-62. Applications: Western Blotting.
- Zheng, W. et al. (2011) Blood 118, 1154-62. Applications: Western Blotting.
- Sakamoto, K. et al. (2012) Lab Invest 92, 688-702. Applications: IHC-P (paraffin).
Have you published research involving the use of our products? If so we'd love to hear about it. Please let us know!
- 7071 Phototope®-HRP Western Blot Detection System, Anti-rabbit IgG, HRP-linked Antibody
- 7074 Anti-rabbit IgG, HRP-linked Antibody
- 7720 Prestained Protein Marker, Broad Range (Premixed Format)
- 7727 Biotinylated Protein Ladder Detection Pack
- 7003 20X LumiGLO® Reagent and 20X Peroxide
- 3447 Notch1 (5B5) Rat mAb
- 2420 Notch2 (8A1) Rabbit mAb
- 3446 Notch3 (8G5) Rat mAb
- 2889 Notch3 Antibody
- 2588 DLL1 Antibody
- 2483 DLL3 (G93) Antibody
- 2589 DLL4 Antibody
- 2155 Jagged1 (1C4) Rabbit mAb
- 2620 Jagged1 (28H8) Rabbit mAb
- 2210 Jagged2 (C23D2) Rabbit mAb
- 2205 Jagged2 (C83A8) Rabbit mAb
- 3608 Notch1 (D1E11) XP® Rabbit mAb
- 2756 Numb (C29G11) Rabbit mAb
- 2761 Numb (C44B4) Rabbit mAb
- 2154 Musashi Antibody
- 3632 Nicastrin Antibody
- 9002 SimpleChIP® Enzymatic Chromatin IP Kit (Agarose Beads)
- 9003 SimpleChIP® Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9004 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Agarose Beads)
- 9005 SimpleChIP® Plus Enzymatic Chromatin IP Kit (Magnetic Beads)
- 9006 ChIP-Grade Protein G Magnetic Beads
- 9007 ChIP-Grade Protein G Agarose Beads
- 7017 6-Tube Magnetic Separation Rack
- 7273 SimpleChIP® Human HES4 Promoter Primers
- 4486 SimpleChIP® Human α Satellite Repeat Primers
- 8216 PhosphoPlus® Notch1 (Cleaved, Val1744) Antibody Duet
For Research Use Only. Not For Use In Diagnostic Procedures.
Cell Signaling Technology® is a trademark of Cell Signaling Technology, Inc.