Notch-1 Antibody - BSA Free

Novus Biologicals | Catalog # NBP1-78292

Novus Biologicals
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Key Product Details

Species Reactivity

Validated:

Human, Mouse

Cited:

Human, Mouse

Applications

Validated:

Immunohistochemistry, Immunohistochemistry-Paraffin, Western Blot, Immunocytochemistry/ Immunofluorescence, Simple Western

Cited:

Western Blot, Immunocytochemistry/ Immunofluorescence, Knockdown Validated

Label

Unconjugated

Antibody Source

Polyclonal Rabbit IgG

Format

BSA Free
View all Notch-1 Primary Antibodies »

Product Specifications

Immunogen

A synthetic peptide made to a C-terminal portion of the human NOTCH1 protein (between residues 2300-2350) [UniProt# P46531]

Localization

Cell membrane; Single-pass type I membrane protein. Notch 1 intracellular domain: Following proteolytical processing is translocated to the nucleus.

Clonality

Polyclonal

Host

Rabbit

Isotype

IgG

Scientific Data Images for Notch-1 Antibody - BSA Free

Western Blot: Notch-1 AntibodyBSA Free [NBP1-78292]

Western Blot: Notch-1 AntibodyBSA Free [NBP1-78292]

Western Blot: Notch-1 Antibody [NBP1-78292] - Mouse total protein lysates (50 ug) were separated on 8% SDS-PAGE. Notch-1 (NBP1-78292SS) antibody was used at 1:500 dilution. Actin (1:10,000) was used as normalizer. WB image submitted by a verified customer review.
Immunohistochemistry: Notch-1 Antibody - BSA Free [NBP1-78292]

Immunohistochemistry: Notch-1 Antibody - BSA Free [NBP1-78292]

Immunohistochemistry: Notch-1 Antibody [NBP1-78292] - Staining of Notch1 in human kidney carcinoma using DAB with hematoxylin counterstain.
Western Blot: Notch-1 AntibodyBSA Free [NBP1-78292]

Western Blot: Notch-1 AntibodyBSA Free [NBP1-78292]

Western Blot: Notch-1 Antibody [NBP1-78292] - Analysis of Cleaved Notch1 in Jurkat cell lysate.
Simple Western: Notch-1 AntibodyBSA Free [NBP1-78292]

Simple Western: Notch-1 AntibodyBSA Free [NBP1-78292]

Simple Western: Notch-1 Antibody [NBP1-78292] - Lane view shows a specific band for Notch-1 in 0.5 mg/ml of Jurkat lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.

Applications for Notch-1 Antibody - BSA Free

Application
Recommended Usage

Immunocytochemistry/ Immunofluorescence

1:10-1:500

Immunohistochemistry

1:200

Immunohistochemistry-Paraffin

1:200

Simple Western

1:50

Western Blot

2 ug/ml
Application Notes
This Notch1 antibody is useful for Western Blot and Immunohistochemistry-Paraffin. In Western Blot, a band is seen ~120 kDa representing the cleaved form of Notch1 peptide. In IHC-P, staining of nucleus and cell membrane was observed in human kidney carcinoma tissue. Prior to immunostaining paraffin tissues, antigen retrieval with sodium citrate buffer (pH 6.0) is recommended.

In Simple Western only 10 - 15 uL of the recommended dilution is used per data point.
See Simple Western Antibody Database for Simple Western validation: Tested in Jurkat lysate 0.5 mg/mL, separated by Size, antibody dilution of 1:50, apparent MW was 125 kDa. Separated by Size-Wes, Sally Sue/Peggy Sue.

Reviewed Applications

Read 1 review rated 5 using NBP1-78292 in the following applications:

Formulation, Preparation, and Storage

Purification

Immunogen affinity purified

Formulation

PBS and 30% Glycerol

Format

BSA Free

Preservative

0.05% Sodium Azide

Concentration

1.18 mg/ml

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & Storage

Store at 4C short term. Aliquot and store at -20C long term. Avoid freeze-thaw cycles.

Background: Notch-1

Notch1 is a single-pass transmembrane protein that serves as the receptor for the Delta1, Jagged1 and Jagged2 ligands. Activation of Notch1 causes the cytoplasmic domain to be cleaved, releasing the notch intracellular domain (NICD) which forms a complex with RBPJ/RBPSUH and subsequently influences transcription of its target genes. Because the majority of Notch1 ligands are transmembrane proteins, Notch1 is primarily triggered by cell-to-cell contact and employs lateral inhibition mechanisms to influence local populations of cells. The Notch1 pathway plays a pivotal role in development, including cell-fate determination, proliferation, and cell contact-dependent signaling. Aberrant activation of the Notch1 pathway has been associated with numerous diseases such as bicuspid aortic valve and cancer-- in particular certain leukemias.

Alternate Names

Notch1, TAN1

Entrez Gene IDs

4851 (Human); 18128 (Mouse)

Gene Symbol

NOTCH1

UniProt

P46531

Additional Notch-1 Products

Product Documents for Notch-1 Antibody - BSA Free

Certificate of Analysis

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

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Product Specific Notices for Notch-1 Antibody - BSA Free

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Citations for Notch-1 Antibody - BSA Free

Customer Reviews for Notch-1 Antibody - BSA Free (1)

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Customer Images

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  • Notch-1 Antibody
    Name: Manjinder Cheema
    Application: Western Blot
    Sample Tested: Mouse brain, mouse liver, Mouse Spleen and Mouse kidney
    Species: Mouse
    Verified Customer | Posted 02/18/2020
    Mouse total protein lysates (50ug) were separated on 8% SDS-PAGE. Notch-1 (NBP1-78292SS) antibody was used at 1:500 dilution. Actin (1:10,000) was used as normalizer.
    Notch-1 Antibody - BSA Free NBP1-78292

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Protocols

View specific protocols for Notch-1 Antibody - BSA Free (NBP1-78292):

Notch-1 Antibody:
Immunohistochemistry-Paraffin Embedded Sections

Antigen Unmasking:
Bring slides to a boil in 10 mM sodium citrate buffer (pH 6.0) then maintain at a sub-boiling temperature for 10 minutes. Cool slides on bench-top for 30 minutes.

Staining:
1. Wash sections in deionized water three times for 5 minutes each.
2. Wash sections in wash buffer for 5 minutes.
3. Block each section with 100-400 ul blocking solution for 1 hour at room temperature.
4. Remove blocking solution and add 100-400 ul diluted primary antibody. Incubate overnight at 4C.
5. Remove antibody solution and wash sections in wash buffer three times for 5 minutes each.
6. Add 100-400 ul biotinylated diluted secondary antibody. Incubate 30 minutes at room temperature.
7. Remove secondary antibody solution and wash sections three times with wash buffer for 5 minutes each.
8. Add 100-400 ul Streptavidin-HRP reagent to each section and incubate for 30 minutes at room temperature.
9. Wash sections three times in wash buffer for 5 minutes each.
10. Add 100-400 ul DAB substrate to each section and monitor staining closely.
11. As soon as the sections develop, immerse slides in deionized water.
12. Counterstain sections in hematoxylin.
13. Wash sections in deionized water two times for 5 minutes each.
14. Dehydrate sections.
15. Mount coverslips.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Notch-1 Antibody:
Western Blot Protocol

1. Perform SDS-PAGE on samples to be analyzed, loading 40 ug of total protein per lane.
2. Transfer proteins to membrane according to the instructions provided by the manufacturer of the membrane and transfer apparatus.
3. Stain according to standard Ponceau S procedure (or similar product) to assess transfer success, and mark molecular weight standards where appropriate.
4. Rinse the blot.
5. Block the membrane using standard blocking buffer for at least 1 hour.
6. Wash the membrane in wash buffer three times for 10 minutes each.
7. Dilute primary antibody in blocking buffer and incubate 1 hour at room temperature.
8. Wash the membrane in wash buffer three times for 10 minutes each.
9. Apply the diluted HRP conjugated secondary antibody in blocking buffer (as per manufacturers instructions) and incubate 1 hour at room temperature.
10. Wash the blot in wash buffer three times for 10 minutes each (this step can be repeated as required to reduce background).
11. Apply the detection reagent of choice in accordance with the manufacturers instructions.
Note: Tween-20 can be added to the blocking or antibody dilution buffer at a final concentration of 0.05-0.2%.

*The above information is only intended as a guide. The researcher should determine what protocol best meets their needs. Please follow safe laboratory procedures.

Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.

FAQs for Notch-1 Antibody - BSA Free

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  • Q: Hello, does this product (NB300-251) only recognize the cleavage Notch1?

    A: This antibody with recognize full length Notch1 if it is in the mix, as well as the cleaved product.

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