Human EPOR & CD131 Heterodimer HEK293 Overexpression Lysate: 製品の情報
This Human EPOR & CD131 Heterodimer overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of EPOR & CD131 Heterodimer protein (Cat: CT010-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Pro 250) of human EPOR (NP_000112.1) linked with the extracellular domain (Trp 17-Trp 443) of human CD131 (NP_000386.1) by a peptide linker was fused with the Fc region of human IgG1 at the C-terminus.
The recombinant human EPOR&CD131/Fc chimera is a disulfide-linked homodimer of EPOR&CD131-Fc heterodimer. The reduced EPOR& CD131-Fc comprises 906 amino acids and has a calculated molecular mass of 101 kDa. As a result of glycosylation, the apparent molecular mass of the recombinant protein is approximately 125 kDa in SDS-PAGE under reducing conditions.
Human EPOR & CD131 Heterodimer HEK293 Overexpression Lysate: 用法
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.