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Encompassing Amino Acids: 19-333
Application: Useful for binding assays and avidin-pull down assays.
Assay Conditions: A white 96-well plate was precoated with ITGA4-ITGB7 protein (AcroBiosystems #IT7-H52W4) (200 ng/well). Superblock™ TBS Blocking Buffer (Thermo Fisher Scientific #37581) with 2 mM MnCl2 and TBST with 2 mM MnCl2 were used for blocking and washing steps, respectively. For protein titration a serial dilution of MAdCAM1, Fc-Fusion (IgG1), Avi-Tag, Biotin-Labeled Recombinant (BPS Bioscience #102495) at the highest concentration of 10 ng/µl (250 ng/reaction) was prepared in 1x Assay Buffer. Diluted MAdCAM1 was then added to the wells and incubated at Room Temperature (RT) for 1 hour. The plate was washed again and incubated with Streptavidin-HRP diluted 1:1000 in Superblock™ TBS Blocking Buffer with 2 mM MnCl2 for 1 hour at RT. After the final wash, ELISA ECL mix (BPS Bioscience #79670) was added to each well and chemiluminescence was read.
Background: MAdCAM1 (Mucosal addressin cell adhesion molecule 1) is a member of the immunoglobulin superfamily involved in guiding memory T cells and other immune cells to mucosal tissues. MAdCAM1 binds to lymphocyte receptor LPAM-1 (lymphocyte Peyer’s patch HEV adhesion molecule 1), also known as integrin α4β7 (ITGA4-ITGB7), a key interaction that guides lymphocytes to the gastrointestinal system. MAdCAM1 is linked to inflammatory bowel diseases, such as Crohn’s disease and ulcerative colitis (UC), as well as colon cancer. MAdCAM1-targeting monoclonal antibodies and small molecule inhibitors aim to alleviate these gastrointestinal diseases, including FDA-approved Vedolizumab, and other therapeutics currently undergoing clinical trials.
Description: Recombinant MAdCAM1 (Mucosal addressin cell adhesion molecule 1), encompassing amino acids 19-333. This construct contains at the C-terminus the Fc domain of a human IgG1, followed by an Avi-Tag™. This protein is enzymatically biotinylated using Avi-Tag™ technology and affinity purified.
Format: Aqueous buffer solution
Formulation: 8 mM phosphate, 110 mM NaCl, 2.2 mM KCl, pH 7.4, and 20% glycerol
Genbank: DQ076657
Host Cell Line: HEK293
Purity: ≥85%
Storage Stability: At least 6 months at -80°C.
Tags: C-terminal Fc (IgG1)-Avi-tag
Target: MAdCAM1
Uniprot: Q4PKD0-1
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
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