Notch2, Fc Fusion, Avi-Tag, Biotin-Labeled Recombinant

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Catalog #

101987

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Description

Recombinant human Notch2 (neurogenic locus notch homolog protein 2), encompassing amino acids 26-530 (extracellular domain). This construct contains a C-terminal Fc domain from IgG1 followed by an Avi-tag™. This protein was affinity purified.

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Synonyms

Neurogenic locus notch homolog protein 2, Notch 2, hN2, Notch 2 extracellular truncation, Notch 2 intracellular domain, NOTCH2

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Product Data Gallery

4-20% SDS-PAGE Coomassie Staining

Biotin-Avidin Pulldown

Anti Notch:Notch2-Biotin Binding Assay

Product Info

Storage and Usage

Citations

Species

Human

Construct

Notch2 (26-530-Fc(lgG1)-Avi)-(Biotin)

Host Species/Expression System

HEK293

Purity

≥90%

Format

Aqueous buffer solution.

Formulation

8 mM phosphate, pH 7.4, 110 mM NaCl, 2.2 mM KCl, and 20% glycerol

83 kDa + glycans

Amino Acids

26-530

Glycosylation

This protein runs at a higher MW by SDS-PAGE due to glycosylation.

Genbank #

NM_024408.4

UniProt #

Q04721

Tag(s)

C-terminal Avi-Tag™

Label

This protein is enzymatically biotinylated using Avi-Tag™ technology. Biotinylation is confirmed to be ≥90%.

For more information on enzymatic biotinylation, please see our Tech Note.

Background

Notch2 (neurogenic locus notch homolog protein 2) is a type I transmembrane protein of the Notch family. It is involved in the development of the vascular network, brain, kidneys, liver, T and B lymphocytes. The protein undergoes cleavage in the trans-golgi network, forming a heterodimer. Binding of Notch2 to its ligands of the Delta/Serrate/LAG-2 family leads to its cleavage by ADAM (A disintegrin and metaloprotease) proteins. NICD (notch intracellular domain) is released and translocated to the nucleus, where it can form complexes with DNA-binding proteins and lead to transcription of target genes. Mutations in this protein can result in Hadju-Cheney syndrome. Overexpression of Notch2 is found in several cancer types, such as LSCC (laryngeal squamous cell carcinoma) and B cell malignancies. The use of inhibitors to target this protein is being studied with the goal of understanding better the role of this protein and potentially target it in cancer therapy.

Storage/Stability

At least 6 months at -80°C.

Assay Conditions

The protein was validated by measuring Notch2 binding to anti-Notch in an ELISA assay. Anti-Notch Antibody (BPS Bioscience #102065) was coated onto a 96-well plate overnight at 4°C (50 µl/well at a concentration of 2 µg/ml in PBS). The plate was washed 3 times with Immuno Buffer 1 (BPS Bioscience #79311) and blocked using 100 µl of Blocking Buffer 2 (BPS Bioscience #79728) for 1 hour at Room Temperature (RT). After removing the blocking buffer, 50 µl/well of serially diluted Notch2, Fc Fusion, Avi-Tag, Biotin-Labeled Recombinant (BPS Bioscience #101987) in Blocking Buffer 2, was added for 1 hour at RT. After 3 washes, the plate was incubated with Streptavidin-HRP (BPS Bioscience #79742). This was followed by washing and incubation with Colorimetric HRP substrate. The reaction was stopped, and absorbance was read at λ= 450 nm. The Blank value was subtracted from all values

Instructions for Use

Thaw on ice and gently mix prior to use. DO NOT VORTEX. Perform a quick spin before opening. Aliquot into small volumes and flash freeze for long term storage. Avoid multiple freeze/thaw cycles.

Applications

Useful for studying the binding of Notch2 in ELISA and in cellular assays.

Shipping Temperature

-80°C

Warnings

Avoid freeze/thaw cycles

Scientific Category

Cell Signaling Pathway

Regulatory Status

Research Use Only