Human Notch-1 Biotinylated Antibody

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BAM53171
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Human Notch-1 Biotinylated Antibody Summary

Species Reactivity
Human
Specificity
Detects human Notch-1 in ELISAs.
Source
Monoclonal Mouse IgG1 Clone # 527422
Purification
Protein A or G purified from hybridoma culture supernatant
Immunogen
Mouse myeloma cell line NS0-derived recombinant human Notch-1
Ala19-Gln526
Accession # P46531
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with BSA as a carrier protein.
Label
Biotin

Applications

Recommended Concentration
Sample

Human Notch-1 Sandwich Immunoassay

Recommended Concentration
Reagent
ELISA Detection (Matched Antibody Pair)
0.5-2.0 µg/mL 

Use in combination with:

Capture Reagent: Human Notch‑1 Antibody (Catalog # MAB5317)

Standard: Recombinant Human Notch-1 Fc Chimera Protein, CF (Catalog # 3647-TK)

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

Reconstitution
Reconstitute at 0.5 mg/mL in sterile PBS.
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Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.

Background: Notch-1

Human Notch-1 is a 300 kDa type I transmembrane glycoprotein that is one of four human 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, promoting T- over B-cell development from a common precursor (2). More than 50% of human T-lineage acute lymphoblastic leukemia (T-ALL) have activating mutations of Notch1 (1, 5). Human Notch-1 is synthesized as a 2556 amino acid (aa) precursor that contains an 18 aa signal sequence, a 1718 aa extracellular domain (ECD) with 36 EGF-like repeats and three Lin-12/Notch repeats (LNR), a 23 aa transmembrane (TM) segment and a 785 aa cytoplasmic domain containing six ankyrin repeats, a glutamine-rich domain and a PEST sequence. The 11th and 12th EGF-like repeats bind ligands including Jagged and Delta-like families in humans (6). O-fucosylation 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). Notch-1 receptor undergoes post-translational furin-type proteolytic cleavage, forming a heterodimer through interaction of a hydrophobic area C-terminal to the LNR on the 1647 aa ligand-binding extracellular region with the 891 aa transmembrane/cytoplasmic portion (8, 9). Upon ligand binding, additional sequential proteolysis by TNF-converting enzyme (ADAM-17) 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). Human Notch-1 ECD aa 19 - 526, including the first 13 EGF repeats, shows 91% aa identity with corresponding regions of mouse and rat, 89% with canine, and 79% with chicken Notch-1. This region also exhibits 60% aa identity with human Notch-2 and Notch-3.

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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