Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody

Name: Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody
Brand: R&D Systems
SKU: FAB5267R

R&D Systems, Inc. a Bio-Techne Brand

Catalog # FAB5267R

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Species Reactivity:	Mouse
Immunogen:	Mouse embryonic fibroblast cell line PA317-derived recombinant mouse Notch-1 Accession # Q01705
Applications:	Flow Cytometry
Label:	Alexa Fluor 647 (Excitation=650 nm, Emission=668 nm)
Source:	Monoclonal Mouse IgG₁ Clone # N1A
Specificity:	Detects mouse Notch-1 intracellular domain in direct ELISAs.

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Applications for Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody

	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.

Formulation, Preparation and Storage

Purification	Protein A or G purified from hybridoma culture supernatant
Formulation	Supplied 0.2 mg/mL in a saline solution containing BSA and Sodium Azide.
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.

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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 11^th and 12^th 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).

Item	Value
References	Ellisen, L.W. et al. (1991) Cell 66:649. Dumortier, A. et al. (2005) Int. J. Hematol. 82:277. Yoon, K. and N. Gaiano (2005) Nat. Neurosci. 8:709. Androutsellis-Theotokis, A. et al. (2006) Nature 442:823. Weng, A.P. et al. (2004) Science 306:269. Hambleton, S. et al. (2004) Structure 12:2173. Yang, L. et al. (2005) Mol. Biol. Cell 16:927. Sanchez-Irizarry, C. et al. (2004) Mol. Cell. Biol. 24:9265. Logeat, F. et al. (1998) Proc. Natl. Acad. Sci. USA 95:8108. 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, Notch (Drosophila) homolog 1 (translocation-associated), Notch homolog 1, translocation-associated, Notch-1, Notch1, TAN1, TAN1neurogenic locus notch homolog protein 1, Translocation-associated notch protein TAN-1, hN1, notch 1Notch homolog 1, translocation-associated (Drosophila)

Product Specific Notices for Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody

This product is provided under an agreement between Life Technologies Corporation and R&D Systems, Inc, and the manufacture, use, sale or import of this product is subject to one or more US patents and corresponding non-US equivalents, owned by Life Technologies Corporation and its affiliates. The purchase of this product conveys to the buyer the non-transferable right to use the purchased amount of the product and components of the product only in research conducted by the buyer (whether the buyer is an academic or for-profit entity). The sale of this product is expressly conditioned on the buyer not using the product or its components (1) in manufacturing; (2) to provide a service, information, or data to an unaffiliated third party for payment; (3) for therapeutic, diagnostic or prophylactic purposes; (4) to resell, sell, or otherwise transfer this product or its components to any third party, or for any other commercial purpose. Life Technologies Corporation will not assert a claim against the buyer of the infringement of the above patents based on the manufacture, use or sale of a commercial product developed in research by the buyer in which this product or its components was employed, provided that neither this product nor any of its components was used in the manufacture of such product. For information on purchasing a license to this product for purposes other than research, contact Life Technologies Corporation, Cell Analysis Business Unit, Business Development, 29851 Willow Creek Road, Eugene, OR 97402, Tel: (541) 465-8300. Fax: (541) 335-0354.

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Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody

R&D Systems, Inc. a Bio-Techne Brand

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Formulation, Preparation and Storage

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Background: Notch-1

Product Specific Notices for Mouse Notch-1 Alexa Fluor® 647-conjugated Antibody

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