Product Pathways - Development
Notch Isoform Antibody Sampler Kit #3640
|3640S||1 Kit (4 x 40 µl)||---||In Stock||---|
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|Kit Includes||Quantity||Applications||Reactivity||MW (kDa)||Isotype|
|Cleaved Notch1 (Val1744) (D3B8) Rabbit mAb #4147||40 µl||W, IP, ChIP||H, M, R||110||Rabbit IgG|
|Notch1 (D6F11) XP® Rabbit mAb #4380||40 µl||W, IF-IC, F||H, M, R||120, 300||Rabbit IgG|
|Notch3 (D11B8) Rabbit mAb #5276||40 µl||W, IP||H||90, 270||Rabbit IgG|
|Notch2 (D76A6) XP® Rabbit mAb #5732||40 µl||W, IP, IF-IC, F||H, M, R||110, 300||Rabbit|
|Anti-rabbit IgG, HRP-linked Antibody #7074||100 µl||Goat|
Applications Key: W=Western Blotting, IP=Immunoprecipitation, ChIP=Chromatin IP, IF-IC=Immunofluorescence (Immunocytochemistry), F=Flow Cytometry
Reactivity Key: H=Human, M=Mouse, R=Rat
Western blot analysis of extracts from various cell lines using Cleaved Notch1 (Val1744) (D3B8) XP® Rabbit mAb #4147 (upper) or Notch1 (D1E11) Rabbit mAb #3608 (lower).
Western blot analysis of extracts from HBP-ALL, and Molt4 using Notch1 (D6F11) XP® Rabbit mAb #4380. The full-length (FL) and transmembrane/intracellular region (NTM) are indicated.
Western blot analysis of extracts from various cell lines using Notch3 (D11B8) XP® Rabbit mAb #5276.
The Notch Isoform Antibody Sampler Kit provides an economical means to investigate Notch Signaling. The kit contains primary and secondary antibodies to perform four western mini-blots with each antibody.
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). Notch1 (D6F11) XP® Rabbit detects endogenous level of total notch1 protein. It recognizes both the full-length (~300 kDa) and the transmembrane/intracellular region NTM (~120 kDa). Notch2 (D76A6) XP® Rabbit mAb detects endogenous levels of total Notch2 protein. It recognizes both the full-length (~ 300 kDa) and the transmembrane/intracellular region NTM (~110 kDa). Notch3 (D11B8) Rabbit mAb detects endogenous levels of total Notch3 protein. The antibody recognizes both full-length (FL) Notch3 at 270 kDa and a truncated protein (NTM) containing a short extracellullar region, the transmembrane domain and the intracellular region at 90 kDa.
Source / Purification
Monoclonal antibodies are 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), Gln2497 of human Notch1, Ala2738 of human Notch2, or Glu2312 of human Notch3 protein.
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).
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 (5-6). Notch2 is a member of the Notch family and mutation in Notch2 is associated with Alagille syndrome (7). Notch3 is a member of the Notch family and is processed similar to Notch1 (8). 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 (9-11). Recent studies indicate that Notch3 is overexpressed in many types of cancer (12-14).
- 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.
- McDaniell, R. et al. (2006) Am J Hum Genet 79, 169-73.
- Baron, M. (2003) Semin Cell Dev Biol 14, 113-9.
- Kalimo, H. et al. (2002) Brain Pathol 12, 371-84.
- Karlström, H. et al. (2002) Proc Natl Acad Sci U S A 99, 17119-24.
- Monet, M. et al. (2007) Hum Mol Genet 16, 982-92.
- Park, J.T. et al. (2006) Cancer Res 66, 6312-8.
- Gramantieri, L. et al. (2007) Liver Int 27, 997-1007.
- Yamaguchi, N. et al. (2008) Cancer Res 68, 1881-8.
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* Product-specific protocol.
For Research Use Only. Not For Use In Diagnostic Procedures.
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