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Application: Useful for studying enzyme kinetics, substrate specificity, and screening inhibitors.
Assay Conditions: Reaction done according to LTBR:LIGHT[B] assay kit (BPS bioscience catalog # 79808) protocol. Buffer is PBS + 0.1% BSA. Blocking buffer is Superblock PBS + 0.05% Tween-20. Coat protein (LTBR) was added overnight at 4 deg 100 ng/well. Various amounts of biotinylated ligand (LIGHT[B]) (0-240 ng) added for 1h at RT. Strep-HRP added for 1h at RT. Luminescence developed with ELISA ECL substrate (BPS Bioscience catalog #79670).
Description: Human LIGHT, also known as TNFSF14, HVEM-L, and LT-y, GenBank Accession No. NM_003807, a.a. 72-240(end) with an N-terminal His-tag and C-terminal Avi-tag, expressed in a HEK293 cell expression system and enzymatically biotin-labeled using Avi-tag™ technology. Biotinylation is confirmed to be ≥90%. MW=23 kDa. This protein runs at a higher molecular weight due to glycosylation.
Formulation: 8 mM phosphate, 110 mM NaCl, 2.2 mM KCl, 20% glycerol.
Genbank: NM_003807
Storage Stability: At least 6 months at -80°C.
Supplied As: Aqueous buffer solution.
Tags: N-terminal His-tag and C-terminal Avi-tag
Uniprot: O43557
Warnings: Avoid freeze/thaw cycles.
Biosafety Level: Not applicable (BSL-1)
References: 1. Edwards, J.R. et al., Arthritis & Rheumatology. 2006; 54(5): 1451-1462. 2. Ogawa, T. et al., The J. or Urology. 2010; 183(3): 1206-1212.