This Mouse CAR/CXADR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CAR/CXADR protein (Cat: 50019-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse CXADR (NP_001020363.1) extracellular domain (Met 1-Gly 237) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant mouse CXADR consists of 229 amino acids and has a predicted molecular mass of 25.7 kDa. As a result of glycosylation, rmCXADR migrates as an approximately 35 kDa band in SDS-PAGE under reducing conditions.
Cell lysate was prepared by homogenization of the over-expressed cells in ice-cold modified RIPA Lysis Buffer with cocktail of protease inhibitors (Sigma). Cell debris was removed by centrifugation. Protein concentration was determined by Bradford assay (Bio-Rad protein assay, Microplate Standard assay). The cell lysate was boiled for 5 min in 1 x SDS loading buffer (50 mM Tris-HCl pH 6.8, 12.5% glycerol, 1% sodium dodecylsulfate, 0.01% bromophenol blue) containing 5% b-mercaptoethanol, and lyophilized.
CXADR (coxsackie virus and adenovirus receptor), also known as CAR, is a type I transmembrane glycoprotein belonging to the CTX family of the Ig superfamily, and is essential for normal cardiac development in the mouse. Proposed as a homophilic cell adhesion molecule, CXADR is a component of the epithelial apical junction complex that is essential for the tight junction integrity, and probably involved in transepithelial migration of polymorphonuclear leukocytes (PMN). Mature mouse CXADR structrually comprises a 218 aa extracellular domain (ECD) with a V-type (D1) and a C2-type (D2) Ig-like domain, a 21 aa transmembrane segment and a 17 aa intracellular domain, among which,D1 is thought to be responsible for homodimer formation in trans within tight junctions. The ECD of mouse CXADR shares 97%, 9% sequence identity with the corresponding regions of rat, human CXADR.
coxsackie virus and adenovirus receptor
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