Mouse CAR HEK293 Overexpression Lysate 产品信息
Product Description
This Mouse CAR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CAR protein (Cat: 50019-M03H1) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the mouse Cxadr (NP_001020363.1) (Met1-Gly237) was expressed with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus (Fc-his).
Molecule Mass
The recombinant mouse Cxadr consists of 465 amino acids and predicts a molecular mass of 52 kDa.
Mouse CAR HEK293 Overexpression Lysate Usage Guide
Preparation Method
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.
Lysis Buffer
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
Recommend Usage
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.
Sample Buffer
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.
Mouse CAR HEK293 Overexpression Lysate Alternative Names
Mouse 2610206D03Rik Overexpression Lysate;Mouse AU016810 Overexpression Lysate;Mouse AW553441 Overexpression Lysate;Mouse CAR Overexpression Lysate;Mouse MCAR Overexpression Lysate;Mouse MCVADR Overexpression Lysate
CAR 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.
Full Name
coxsackie virus and adenovirus receptor
References
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. Carson, S.D. et al., 2002, Rev. Med. Virol. 11 (4): 219–226. Selinka, H.C. et al., 2004, Med. Microbiol. Immunol. 193 (2-3): 127–131. Philipson, L. et al., 2004, Curr. Top. Microbiol. Immunol. 273:87-111. Raschperger, E. et al., 2006, Exp. Cell Res. 312: 1566-1580.