Mouse Notch-1 Antibody

Name: Mouse Notch-1 Antibody
Brand: R&D Systems
SKU: AF5267

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

Catalog # AF5267

Citations (7)

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Species Reactivity:	Mouse
Immunogen:	Chinese hamster ovary cell line CHO-derived recombinant mouse Notch‑1 Ala19-Gln526 Accession # AAM28905
Applications:	Immunohistochemistry, Western Blot
Label:	Unconjugated
Source:	Polyclonal Sheep IgG
Specificity:	Detects mouse Notch-1 in direct ELISAs and Western blots. In direct ELISA, approximately 10% cross-reactivity with recombinant human Notch-1 and recombinant rat Notch-1 is observed and less than 1% cross-reactivity with recombinant mouse (rm) Notch-2 and rmNotch-3 is observed.

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Applications for Mouse Notch-1 Antibody

	Recommended Concentration	Sample
Western Blot	1 µg/mL	See below
Immunohistochemistry	5-15 µg/mL	See below

Please Note: Optimal dilutions should be determined by each laboratory for each application.

Scientific Data Examples for Mouse Notch-1 Antibody

	Western Blot Detection of Mouse Notch‑1 by Western Blot. Western blot shows lysates of bEnd.3 mouse endothelioma cell line. PVDF membrane was probed with 1 µg/mL of Mouse Notch-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5267) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Notch-1 at approximately 300 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 8.
	Immunohistochemistry Notch‑1 in Mouse Neural Tube. Notch-1 was detected in immersion fixed frozen sections of mouse neural tube (E13.5) using Mouse Notch-1 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF5267) at 10 µg/mL overnight at 4 °C. Tissue was stained using the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red, upper panel; Catalog # NL010) and counterstained with DAPI (blue, lower panel). View our protocol for Fluorescent IHC Staining of Frozen Tissue Sections.

Formulation, Preparation and Storage

Purification	Antigen Affinity-purified
Reconstitution	Reconstitute at 0.2 mg/mL in sterile PBS. Reconstitution Buffer Available: Size / Price Qty Loading... RB01:
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.
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.

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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). Mouse Notch-1 ECD aa 19‑526, which includes the first 13 EGF repeats, shows 94%, 91%, 86% and 79% aa identity with corresponding regions of rat, human, canine, and chicken Notch-1, respectively. This region also exhibits 55‑58% aa identity with human Notch-2 and Notch-3.

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)

Citations for Mouse Notch-1 Antibody (7)

Showing 1 - 7 of 7 citations Showing All

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Citations are publications that use Bio-Techne products. Selected citations for Mouse Notch-1 Antibody include:

Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry

K Matsumoto et al. (2022), Fringe GlcNAc-transferases differentially extend O-fucose on endogenous NOTCH1 in mouse activated T cells The Journal of Biological Chemistry, 0(0):102064.
PMID: 35623385
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry

S Kakuda et al. (2020), Canonical Notch ligands and Fringes have distinct effects on NOTCH1 and NOTCH2 J. Biol. Chem., 0(0).
PMID: 32820046
Species: Mouse
Sample Types: Cell Lysates
Applications: Western Blot

S Kakuda et al. (2020), Canonical Notch ligands and Fringes have distinct effects on NOTCH1 and NOTCH2 J. Biol. Chem., 0(0).
PMID: 32820046
Species: Mouse
Sample Types: Whole Tissue
Applications: IHC-F

S Wiszniak et al. (2019), Notch signalling defines dorsal root ganglia neuroglial fate choice during early neural crest cell migration BMC Neurosci, 20(1):21.
PMID: 31036074
Species: Mouse
Sample Types: Whole Cells
Applications: Flow Cytometry

H Takeuchi et al. (2018), Two novel protein O-glucosyltransferases that modify sites distinct from POGLUT1 and affect Notch trafficking and signaling Proc. Natl. Acad. Sci. U.S.A., 0(0).
PMID: 30127001
Species: Hamster
Sample Types: Whole Cells
Applications: Flow Cytometry

Y Wang et al. (2017), Uncontrolled angiogenic precursor expansion causes coronary artery anomalies in mice lacking Pofut1 Nat Commun, 8(1):578.
PMID: 28924218
Species: Hamster
Sample Types: Cell Lysates
Applications: Western Blot

Tashima Y et al. (2014), Antibodies that detect O-linked beta-D-N-acetylglucosamine on the extracellular domain of cell surface glycoproteins. J Biol Chem, 289(16):11132-42.
PMID: 24573683

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Showing 1 - 7 of 7 citations Showing All

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Mouse Notch-1 Antibody

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Background: Notch-1

Citations for Mouse Notch-1 Antibody (7)

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