Anti-Notch1 Antibody (56544)

Host Quantity Applications Species Reactivity Data Sheet  
Mouse 100ug WB,ICC/IF Mouse, Rat
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SKU: 56544 Categories: , ,
Overview
Product Name Anti-Notch1 Antibody (56544)
Description Anti-Notch1 Mouse Monoclonal Antibody
Target Notch1
Species Reactivity Mouse, Rat
Applications WB,ICC/IF
Host Mouse
Clonality Monoclonal
Clone ID S253-32
Isotype IgG1
Immunogen Fusion protein corresponding to aa 20-216 (extracellular N-terminus, EGF- like domains 1-5) of mouse Notch1 (accession no. Q01705).
Properties
Form Liquid
Concentration Lot Specific
Formulation PBS, pH 7.4; 50% glycerol, 0.09% sodium azide.
Buffer Formulation Phosphate Buffered Saline
Buffer pH pH 7.4
Buffer Anti-Microbial 0.09% Sodium Azide
Buffer Cryopreservative 50% Glycerol
Format Purified
Purification Purified by Protein G affinity chromatography
Specificity Information
Specificity This antibody recognizes mouse and rat Notch1. It does not cross-react with Notch2 or Notch3.
Target Name Neurogenic locus notch homolog protein 1
Target ID Notch1
Uniprot ID Q01705
Alternative Names Notch 1, Motch A, mT14, p300 [Cleaved into: Notch 1 extracellular truncation
Gene Name Notch1
Sequence Location Cell membrane
Biological Function Functions as a receptor for membrane-bound ligands Jagged-1 (JAG1), Jagged-2 (JAG2) and Delta-1 (DLL1) to regulate cell-fate determination. Upon ligand activation through the released notch intracellular domain (NICD) it forms a transcriptional activator complex with RBPJ/RBPSUH and activates genes of the enhancer of split locus. Affects the implementation of differentiation, proliferation and apoptotic programs. Involved in angiogenesis; negatively regulates endothelial cell proliferation and migration and angiogenic sprouting. Involved in the maturation of both CD4(+) and CD8(+) cells in the thymus. Important for follicular differentiation and possibly cell fate selection within the follicle. During cerebellar development, functions as a receptor for neuronal DNER and is involved in the differentiation of Bergmann glia. Represses neuronal and myogenic differentiation. May play an essential role in postimplantation development, probably in some aspect of cell specification and/or differentiation. May be involved in mesoderm development, somite formation and neurogenesis. May enhance HIF1A function by sequestering HIF1AN away from HIF1A. Required for the THBS4 function in regulating protective astrogenesis from the subventricular zone (SVZ) niche after injury. Involved in determination of left/right symmetry by modulating the balance between motile and immotile (sensory) cilia at the left-right organiser (LRO). {PubMed:15965470, PubMed:18299578, PubMed:23160044, PubMed:23615612}.
Research Areas Neuroscience
Background Members of the Notch trans-membrane protein family share structural characteristics including an extracellular domain consisting of multiple epidermal growth factor-like (EGF) repeats, and an intracellular domain consisting of multiple, different domain types. The Notch signaling network is an intercellular signaling pathway that regulates interactions between physically adjacent cells. The Notch1 protein is cleaved in the trans-Golgi network, and presented on the cell surface as a heterodimer. It functions as a receptor for membrane bound ligands, and may play multiple roles during development. There is evidence that activated Notch 1 and Notch 3 promote differentiation of progenitor cells into astroglia. Notch 1, activated before birth, induces radial glia differentiation,but postnatally induces the differentiation into astrocytes.
Application Images
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Notch1 Monoclonal Antibody, Clone S253-32 (56544). Tissue: Neuroblastoma cells (SH-SY5Y). Species: Human. Fixation: 4% PFA for 15 min. Primary Antibody: Mouse Anti-Notch1 Monoclonal Antibody (56544) at 1:50 for overnight at 4°C with slow rocking. Secondary Antibody: AlexaFluor 488 at 1:1000 for 1 hour at RT. Counterstain: Phalloidin-iFluor 647 (red) F-Actin stain; Hoechst (blue) nuclear stain at 1:800, 1.6mM for 20 min at RT. (A) Hoechst (blue) nuclear stain. (B) Phalloidin-iFluor 647 (red) F-Actin stain. (C) Notch1 Antibody (D) Composite.
Description Western Blot analysis of Rat Brain Membrane showing detection of ~270 kDa Notch1 protein using Mouse Anti-Notch1 Monoclonal Antibody, Clone S253-32 (56544). Lane 1: MW Ladder. Lane 2: Rat Brain Membrane (10 µg). . Load: 10 µg. Block: 5% milk. Primary Antibody: Mouse Anti-Notch1 Monoclonal Antibody (56544) at 1:1000 for 1 hour at RT. Secondary Antibody: Goat Anti-Mouse IgG: HRP at 1:200 for 1 hour at RT. Color Development: TMB solution for 10 min at RT. Predicted/Observed Size: ~270 kDa.
Description Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Notch1 Monoclonal Antibody, Clone S253-32 (56544). Tissue: Neuroblastoma cell line (SK-N-BE). Species: Human. Fixation: 4% Formaldehyde for 15 min at RT. Primary Antibody: Mouse Anti-Notch1 Monoclonal Antibody (56544) at 1:100 for 60 min at RT. Secondary Antibody: Goat Anti-Mouse ATTO 488 at 1:100 for 60 min at RT. Counterstain: Phalloidin Texas Red F-Actin stain; DAPI (blue) nuclear stain at 1:1000, 1:5000 for 60min RT, 5min RT. Localization: Cell Membrane, Nucleus. Magnification: 60X. (A) DAPI (blue) nuclear stain. (B) Phalloidin Texas Red F-Actin stain. (C) Notch1 Antibody. (D) Composite.
Additional Information
Handling
Storage This antibody is stable for at least one (1) year at -20°C.
Dilution Instructions Dilute in PBS or medium that is identical to that used in the assay system.
Application Instructions Immunoblotting: use at 1ug/mL. Bands of >270kDa, ~120kDa, and smaller fragments (due to proteolysis) are detected .

Positive control: Rat brain lysate.

These are recommended concentrations. User should determine optimal concentrations for their application.
References & Data Sheet