Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody

Catalog # Availability Size / Price Qty
FAB5267R-100UG
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Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody Summary

Species Reactivity
Mouse
Specificity
Detects mouse Notch-1 intracellular domain in direct ELISAs.
Source
Monoclonal Mouse IgG1 Clone # N1A
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse embryonic fibroblast cell line PA317-derived recombinant mouse Notch-1
Accession # Q01705
Formulation
Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
Label
Alexa Fluor 647 (Excitation= 650 nm, Emission= 668 nm)

Applications

Recommended Concentration
Sample
Intracellular Staining by Flow Cytometry
0.25-1 µg/106 cells
RPMI 8226 human multiple myeloma cell line fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005)

Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.

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Preparation and Storage

Shipping
The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Protect from light. Do not freeze.
  • 12 months from date of receipt, 2 to 8 °C as supplied.

Background: Notch-1

Notch-1 is a 300 kDa type I transmembrane glycoprotein that is one of four Notch homologues involved in developmental processes (1-3). Notch signaling is important for maintaining stem cells and inducing differentiation, especially in the nervous system and lymphoid tissues (2-4). Notch can specify binary cell fates. For example, it promotes T cell over B cell development from a common precursor (2). Mouse Notch-1 is synthesized as a 2531 amino acid (aa) precursor that contains an 18 aa signal sequence, a 1707 aa extracellular domain (ECD) with 36 EGF-like repeats and three Lin-12/notch repeats (LNR), a 21 aa transmembrane (TM) segment and a 785 aa cytoplasmic domain that contains six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The 11th and 12th EGF-like repeats, that bind ligands such as Jagged and Delta-like families in humans, correspond to aa 412-488 in mouse Notch-1 (6). Elongation of O-linked fucose chains by Fringe family members at a site within this region can inhibit the interaction of Notch with Jagged ligands, thereby promoting Delta-like ligand interactions (7). The Notch-1 receptor undergoes post-translational furin-type proteolytic cleavage, generating a heterodimer through the interaction of a hydrophobic area C-terminal to the LNR on the extracellular region with the transmembrane/cytoplasmic portion (8, 9). Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme (ADAM17) and the presenilin-dependent gamma -secretase results in the release of the Notch intracellular domain (NICD) which translocates into the nucleus, activating transcription of Notch-responsive genes (10).

References
  1. Ellisen, L.W. et al. (1991) Cell 66:649.
  2. Dumortier, A. et al. (2005) Int. J. Hematol. 82:277.
  3. Yoon, K. and N. Gaiano (2005) Nat. Neurosci. 8:709.
  4. Androutsellis-Theotokis, A. et al. (2006) Nature 442:823.
  5. Weng, A.P. et al. (2004) Science 306:269.
  6. Hambleton, S. et al. (2004) Structure 12:2173.
  7. Yang, L. et al. (2005) Mol. Biol. Cell 16:927.
  8. Sanchez-Irizarry, C. et al. (2004) Mol. Cell. Biol. 24:9265.
  9. Logeat, F. et al. (1998) Proc. Natl. Acad. Sci. USA 95:8108.
  10. Mumm, J.S. and R. Kopan (2000) Dev. Biol. 228:151.
Entrez Gene IDs
4851 (Human); 25496 (Rat)
Alternate Names
EC 2.1.2.11; EC 3.4.21.68; hN1; Notch (Drosophila) homolog 1 (translocation-associated); notch 1Notch homolog 1, translocation-associated (Drosophila); Notch homolog 1, translocation-associated; Notch1; Notch-1; TAN1; TAN1neurogenic locus notch homolog protein 1; Translocation-associated notch protein TAN-1

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