Human Notch-1 Intracellular Domain Antibody Summary
Gly2428-Lys2556
Accession # P46531
Applications
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
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Detection of Human Notch-1. Western blot shows lysates of MOLT-4 human acute lymphoblastic leukemia cell line, Jurkat human acute T cell leukemia cell line, RPMI 8226 human multiple myeloma cell line, 293T human embryonic kidney cell line, and 293T human embryonic kidney cell line (1 µg per lane), transfected with full length human Notch-1. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Notch-1 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3647) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for Notch-1 intracellular domain (NICD) and full length Notch-1 (Notch-1 FL) at approximately 115 and 300 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
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Notch‑1 in Saos-2 Human Cell Line. Notch-1 was detected in immersion fixed Saos-2 human osteosarcoma cell line using 10 µg/mL Sheep Anti-Human Notch-1 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3647) for 3 hours at room temperature. Cells were stained with the NorthernLights™ 557-conjugated Anti-Sheep IgG Secondary Antibody (red; Catalog # NL010) and counterstained with DAPI (blue). View our protocol for Fluorescent ICC Staining of Cells on Coverslips.
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Detection of Notch‑1-regulated Genes by Chromatin Immunoprecipitation. Jurkat human acute T cell leukemia cell line treated with 50 ng/mL PMA and 200 ng/mL calcium ionomycin for 30 minutes was fixed using formaldehyde, resuspended in lysis buffer, and sonicated to shear chromatin. Notch-1/DNA complexes were immunoprecipitated using 5 µg Sheep Anti-Human Notch-1 Intracellular Domain Antigen Affinity-purified Polyclonal Antibody (Catalog # AF3647) or control antibody (Catalog # 5-001-A) for 15 minutes in an ultrasonic bath, followed by Biotinylated Anti-Sheep IgG Secondary Antibody (Catalog # BAF016). Immunocomplexes were captured using 50 µL of MagCellect Streptavidin Ferrofluid (Catalog # MAG999) and DNA was purified using chelating resin solution. Thec-mycpromoter was detected by standard PCR.
Reconstitution Calculator
Preparation and Storage
- 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
Notch-1 (so named for "notches" in fly wings; also TAN-1) is a 300 kDa member of the Notch family of glycoproteins. It is associated with gene activation in both embryo and adult. Human Notch-1 is a 2538 amino acid (aa) type I transmembrane glycoprotein. It undergoes Golgi processing to generate a heterodimer composed of a 180‑200 kDa disulfide-linked extracellular domain (aa 18‑1664) and a 120 kDa membrane-bound segment (aa 1665‑2556). Upon ligand binding, the 110 kDa segment undergoes two cleavages which generate an NICD (notch intracellular domain) (aa 1754‑2556), a nuclear transcription factor. One isoform shows a deletion of aa 248‑288. Over aa 2428‑2556, human Notch 1 is 83% and 89% aa identical to canine and mouse Notch-1, respectively.
Product Datasheets
Citations 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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Citations: Showing 1 - 10
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HIV–host interactome revealed directly from infected cells
Authors: Yang Luo, Erica Y. Jacobs, Todd M. Greco, Kevin D. Mohammed, Tommy Tong, Sarah Keegan et al.
Nature Microbiology
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Image-guided genomics of phenotypically heterogeneous populations reveals vascular signalling during symbiotic collective cancer invasion
Authors: J. Konen, E. Summerbell, B. Dwivedi, K. Galior, Y. Hou, L. Rusnak et al.
Nature Communications
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A Notch positive feedback in the intestinal stem cell niche is essential for stem cell self‐renewal
Authors: Kai‐Yuan Chen, Tara Srinivasan, Kuei‐Ling Tung, Julio M Belmonte, Lihua Wang, Preetish Kadur Lakshminarasimha Murthy et al.
Molecular Systems Biology
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Generation of pulmonary neuroendocrine cells and SCLC-like tumors from human embryonic stem cells
Authors: Huanhuan Joyce Chen, Asaf Poran, Arun M. Unni, Sarah Xuelian Huang, Olivier Elemento, Hans-Willem Snoeck et al.
Journal of Experimental Medicine
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AMPA-Kainate Receptor Inhibition Promotes Neurologic Recovery in Premature Rabbits with Intraventricular Hemorrhage
Authors: Praveen Ballabh
J. Neurosci., 2016-03-16;36(11):3363-77.
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Somatic mutations reveal hyperactive Notch signaling and racial disparities in prurigo nodularis
Authors: Rajeh, A;Cornman, HL;Gupta, A;Szeto, MD;Kambala, A;Oladipo, O;Parthasarathy, V;Deng, J;Wheelan, S;Pritchard, T;Kwatra, MM;Semenov, YR;Gusev, A;Yegnasubramanian, S;Kwatra, SG;
medRxiv : the preprint server for health sciences
Species: Human
Sample Types: Whole Tissue
Applications: IHC -
NOTCH1-dependent nitric oxide signaling deficiency in hypoplastic left heart syndrome revealed through patient-specific phenotypes detected in bioengineered cardiogenesis
Authors: SC Hrstka, X Li, TJ Nelson
Stem Cells, 2017-03-05;0(0):.
Species: Human
Sample Types: Cell Lysates, Whole Cells
Applications: ICC, Western Blot -
A dual molecular analogue tuner for dissecting protein function in mammalian cells
Nat Commun, 2016-05-27;7(0):11742.
Species: Human
Sample Types: Cell Lysates
Applications: Western Blot -
Involvement of Notch in activation and effector functions of gammadelta T cells.
Authors: Gogoi D, Dar A, Chiplunkar S
J Immunol, 2014-01-31;192(5):2054-62.
Species: Human
Sample Types: Whole Cells
Applications: Flow Cytometry -
Delta-like 4 inhibits choroidal neovascularization despite opposing effects on vascular endothelium and macrophages
Authors: Serge Camelo, William Raoul, Sophie Lavalette, Bertrand Calippe, Brunella Cristofaro, Olivier Levy et al.
Angiogenesis
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Fixed 4% PFA overnight.
Blocked with 1% BSA
Primary antibody dilution - 1:20
Secondary antibody - Invitrogen Alexa Fluor 488
Secondary antibody dilution - 1:1000
Stained on an E12.5 mouse right ventricle heart section
Expression of cleaved Notch-1 (Intracellular Domain) in mammary tumor cell lines. Dilution: 1:200 in PBS with 5% BSA. Secondary Ab: anti-Sheep IgG 1:5,000.
