Mouse CD51 & ITGB6 Heterodimer HEK293 Overexpression Lysate 产品信息
Product Description
This Mouse CD51 & ITGB6 Heterodimer overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD51 & ITGB6 Heterodimer protein (Cat: CT051-M2508H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Val988) of mouse ITGAV (NP_032428.2) was fused with a flag tag at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the extracellular domain (Met1-Pro708) of mouse ITGB6 (NP_067334.1) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 2. The two plasmids were co-expressed and the mouse ITGAV&ITGB6 heterodimer was purified.
Molecule Mass
The recombinant heterodimer of mouse ITGAV&ITGB6 comprises 1728 (1000+728) amino acids and has a calculated molecular mass of 190.4 (110.7+79.7) KDa.
Mouse CD51 & ITGB6 Heterodimer 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 CD51 & ITGB6 Heterodimer HEK293 Overexpression Lysate Alternative Names
Mouse ITGAV & ITGB6 Overexpression Lysate
CD51 & ITGB6 Heterodimer Background Information
Integrin alpha-5, also known as ITGA5, is a single-pass type I membrane protein which belongs to the integrin alpha chain family. ITGA5 contains 7 FG-GAP repeats. Alpha chain 5 undergoes post-translational cleavage in the extracellular domain to yield disulfide-linked light and heavy chains that join with beta 1 to form a fibronectin receptor. ITGAV&ITGB6 is a receptor for fibronectin and cytotactin. It recognizes the sequence R-G-D in its ligands. Internalisation of ITGAV&ITGB6 via clathrin-mediated endocytosis promotes carcinoma cell invasion.
Full Name
integrin, alpha 5 (fibronectin receptor, alpha polypeptide)
References
Suzuki S., et al.,(1987), Amino acid sequence of the vitronectin receptor alpha subunit and comparative expression of adhesion receptor mRNAs. J. Biol. Chem. 262:14080-14085. Sims M.A., et al., (2000), Cloning and characterisation of ITGAV, the genomic sequence for human cell adhesion protein (vitronectin) receptor alpha subunit, CD51.Cytogenet. Cell Genet. 89:268-271. Ota T., et al.,(2004), Complete sequencing and characterization of 21,243 full-length human cDNAs.Nat. Genet. 36:40-45.
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