Human Notch-1 Intracellular Domain Antibody

Catalog #: MAB3647
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Catalog #	Availability	Size / Price	Qty
MAB3647
MAB3647-SP

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Human Notch-1 Intracellular Domain Antibody Summary

Species Reactivity

Human

Specificity

Detects human Notch-1 Intracellular Domain in Western blots. In Western blots, this antibody does not cross-react with the corresponding intracellular segments of recombinant human (rh) Notch-2, rhNotch-3, or rhNotch-4.

Source

Monoclonal Mouse IgG_2A Clone # 433802

Purification

Protein A or G purified from hybridoma culture supernatant

Immunogen

E. coli-derived recombinant human Notch‑1
Gly2428-Lys2556
Accession # AAG33848

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.

Label

Unconjugated

Applications

Recommended Concentration

Sample

Western Blot

1 µg/mL

Recombinant Human Notch‑1 Intracellular Domain

Immunocytochemistry

8-25 µg/mL

Immersion fixed MOLT‑4 human acute lymphoblastic leukemia cell line

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

Immunocytochemistry

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Notch‑1 in MOLT‑4 Human Cell Line. Notch‑1 was detected in immersion fixed MOLT‑4 human acute lymphoblastic leukemia cell line using Mouse Anti-Human Notch‑1 Intracellular Domain Monoclonal Antibody (Catalog # MAB3647) at 8 µg/mL for 3 hours at room temperature. Cells were stained using the NorthernLights™ 557-conjugated Anti-Mouse IgG Secondary Antibody (red; NL007) and counterstained with DAPI (blue). Specific staining was localized to cell surface and cell nuclei. Staining was performed using our protocol for Fluorescent ICC Staining of Non-adherent Cells.

Reconstitution Calculator

Preparation and Storage

Reconstitution

Reconstitute at 0.5 mg/mL in sterile PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store 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 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 (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). 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 Intracellular Domain 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.

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