Human Notch-1 Intracellular Domain Antibody

Catalog # Availability Size / Price Qty
AF3647
AF3647-SP
Human Notch-1 Intracellular Domain Antibody in Data
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Product Details
Citations (3)
FAQs
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Reviews (3)

Human Notch-1 Intracellular Domain Antibody Summary

Species Reactivity
Human
Specificity
Detects the intracellular domain (ICD) of human Notch-1 in direct ELISAs and Western blots. In direct ELISAs and Western blots,  less than 1% cross-reactivity with recombinant human (rh) Notch-2 ICD, rhNotch-3 ICD, and rhNotch-4 ICD is observed.
Source
Polyclonal Sheep IgG
Purification
Antigen Affinity-purified
Immunogen
E. coli-derived recombinant human Notch‑1
Gly2428-Lys2556
Accession # P46531
Formulation
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.

Applications

Recommended Concentration
Sample
Western Blot
1 µg/mL
See below
Flow Cytometry
2.5 µg/106 cells
U2OS human osteosarcoma cell line
Chromatin Immunoprecipitation (ChIP)
5 µg/5 x 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.
 
Immunocytochemistry
5-15 µg/mL
See below

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.

Data Examples

Western Blot Detection of Human Notch-1. View Larger

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.

Chromatin Immunoprecipitation (ChIP) Detection of Notch‑1-regulated Genes by Chromatin Immunoprecipitation. View Larger

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.

Immunocytochemistry Notch‑1 in Saos-2 Human Cell Line. View Larger

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.

Reconstitution Calculator

Reconstitution Calculator

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Preparation and Storage

Reconstitution
Reconstitute at 0.2 mg/mL in sterile PBS.
Reconstitution Buffer Available
Reconstitution Buffer 1 (PBS)
Catalog #
Availability
Size / Price
Qty
RB01
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

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.

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

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.

3 Citations: Showing 1 - 3
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  1. 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;0(0):.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  2. A dual molecular analogue tuner for dissecting protein function in mammalian cells
    Nat Commun, 2016;7(0):11742.
    Species: Human
    Sample Types: Cell Lysates
    Applications: Western Blot
  3. Involvement of Notch in activation and effector functions of gammadelta T cells.
    Authors: Gogoi D, Dar A, Chiplunkar S
    J Immunol, 2014;192(5):2054-62.

FAQs

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Reviews for Human Notch-1 Intracellular Domain Antibody

Average Rating: 3.7 (Based on 3 Reviews)

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Human Notch-1 Intracellular Domain Antibody
By Anonymous on 01/25/2018
Application: WB Sample Tested: iPS2 human induced pluripotent stem cells Species: Human

Human Notch-1 Intracellular Domain Antibody
By Anonymous on 12/10/2017
Application: ICC/IF Sample Tested: Bladder tissue,Bladder cancer tissue Species: Human

Human Notch-1 Intracellular Domain Antibody
By Anonymous on 03/01/2017
Application: WB Sample Tested: Breast cancer cells Species: Human

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.