Key Product Details

Validated by

Biological Validation

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Western Blot, Flow Cytometry, Immunocytochemistry, Chromatin Immunoprecipitation (ChIP), CyTOF-ready

Cited:

Immunohistochemistry, Western Blot, Flow Cytometry, Immunocytochemistry

Label

Unconjugated

Antibody Source

Polyclonal Sheep IgG
View all Notch-1 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Notch‑1
Gly2428-Lys2556
Accession # P46531

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.

Clonality

Polyclonal

Host

Sheep

Isotype

IgG

Scientific Data Images for Human Notch‑1 Intracellular Domain Antibody

Detection of Human Notch-1.

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.
Notch‑1 antibody in Saos-2 Human Cell Line by Immunocytochemistry (ICC).

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.
Detection of Notch‑1-regulated Genes antibody by Chromatin Immunoprecipitation.

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.

Applications for Human Notch‑1 Intracellular Domain Antibody

Application
Recommended Usage

Chromatin Immunoprecipitation (ChIP)

5 µg/5 x 106 cells
Sample: PMA and calcium ionomycin treated Jurkat human acute T cell leukemia cell line chromatin, c-myc promoter detected by standard PCR

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Flow Cytometry

2.5 µg/106 cells
Sample: U2OS human osteosarcoma cell line

Immunocytochemistry

5-15 µg/mL
Sample: Immersion fixed Saos-2 human osteosarcoma cell line

Western Blot

1 µg/mL
Sample: 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, transfected with full length human Notch-1

Reviewed Applications

Read 4 reviews rated 3.8 using AF3647 in the following applications:

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Flow Cytometry

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

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

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.

Calculators

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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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.

Alternate Names

Notch1, TAN1

Entrez Gene IDs

4851 (Human); 25496 (Rat)

Gene Symbol

NOTCH1

UniProt

P46531

Additional Notch-1 Products

Product Documents for Human Notch‑1 Intracellular Domain Antibody

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

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Product Specific Notices for Human Notch‑1 Intracellular Domain Antibody

For research use only

Citations for Human Notch‑1 Intracellular Domain Antibody

Customer Reviews for Human Notch‑1 Intracellular Domain Antibody (4)

3.8 out of 5
4 Customer Ratings
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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human Notch-1 Intracellular Domain Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: E12.5 mouse embryo fixed in 4% PFA
    Species: Mouse
    Verified Customer | Posted 06/23/2021
    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
    Human Notch‑1 Intracellular Domain Antibody AF3647
  • Human Notch-1 Intracellular Domain Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: iPS2 human induced pluripotent stem cells
    Species: Human
    Verified Customer | Posted 01/25/2018
    Human Notch‑1 Intracellular Domain Antibody AF3647
  • Human Notch-1 Intracellular Domain Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Bladder tissue and Bladder cancer tissue
    Species: Human
    Verified Customer | Posted 12/10/2017
    Human Notch‑1 Intracellular Domain Antibody AF3647
  • Human Notch-1 Intracellular Domain Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Breast cancer cells
    Species: Human
    Verified Customer | Posted 03/01/2017
    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.
    Human Notch‑1 Intracellular Domain Antibody AF3647

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