Human Notch-1 Antibody

Catalog #: MAB5317
1 Citation Datasheet / COA / SDS

Catalog #	Availability	Size / Price	Qty
MAB5317-100
MAB5317-500
MAB5317-SP

Detection of Notch-1 in U2OS Human Cell Line by Flow Cytometry.

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All Notch-1 Antibodies

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Summary Data Examples Preparation and Storage Reconstitution Calculator Background Product Datasheets Related Research Areas

Human Notch-1 Antibody Summary

Species Reactivity

Human

Specificity

Detects human Notch-1.

Source

Monoclonal Mouse IgG₁ Clone # 527425

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

Mouse myeloma cell line NS0-derived recombinant human Notch-1
aa 19-526
Accession # P46531

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Purity

Protein A or G purified from hybridoma culture supernatant

Applications

Recommended Concentration

Sample

Flow Cytometry

2.5 µg/10⁶ cells

See below

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Human Notch-1 Sandwich Immunoassay

Recommended Concentration

Reagent

ELISA Capture (Matched Antibody Pair)

2-8 µg/mL

Use in combination with:

Detection Reagent: Human Notch‑1 Biotinylated Antibody (Catalog # BAM53171)

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.

Scientific Data

Flow Cytometry

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Detection of Notch-1 in U2OS Human Cell Line by Flow Cytometry. U2OS human osteosarcoma cell line was stained with Human Notch‑1 Monoclonal Antibody (Catalog # MAB5317, filled histogram) or isotype control antibody (Catalog # MAB002, open histogram), followed by Allophycocyanin-conjugated Anti-Mouse IgG F(ab')₂Secondary Antibody (Catalog # F0101B).

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C

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 11^th and 12^th 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

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; 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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Citation for Human Notch-1 Antibody

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citation: Showing 1 - 1

The CD46-Jagged1 interaction is critical for human TH1 immunity.
Authors: Le Friec G, Sheppard D, Whiteman P, Karsten C, Shamoun S, Laing A, Bugeon L, Dallman M, Melchionna T, Chillakuri C, Smith R, Drouet C, Couzi L, Fremeaux-Bacchi V, Kohl J, Waddington S, McDonnell J, Baker A, Handford P, Lea S, Kemper C
Nat Immunol, 2012-10-21;13(12):1213-21.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry