Human CAR/CXADR HEK293 Overexpression Lysate: Product Information
This Human 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: 10799-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the human CXADR (NP_001329.1) extracellular domain (Met 1-Gly 237) was fused with a polyhistidine tag at the C-terminus.
The secreted recombinant human CXADR consists of 229 amino acids and has a predicted molecular mass of 25.6 kDa. As a result of glycosylation, the apparent molecular mass of rh CXADR is approximately 35 kDa in SDS-PAGE under reducing conditions.
Human CAR/CXADR HEK293 Overexpression Lysate: Usage Guide
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.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human CAR/CXADR HEK293 Overexpression Lysate: Alternative Names
Human CAR Overexpression Lysate; Human CAR4/6 Overexpression Lysate; Human CXADR Overexpression Lysate; Human HCAR Overexpression Lysate
CAR/CXADR Background Information
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
Tomko, R.P. et al., 1997, Proc. Natl. Acad. Sci. U.S.A. 94 (7): 3352–3356.
van Raaij , M.J. et al., 2001, Structure. 8 (11): 1147–1155.
Cohen, C.J. et al., 2001, J. Biol. Chem. 276 (27): 25392–25398.
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