Human CD1B & B2M Heterodimer HEK293 Overexpression Lysate: 产品信息
This Human CD1B & B2M Heterodimer overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD1B & B2M Heterodimer protein (Cat: CT044-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met1-Ser303) of human CD1B (NP_001755.1) was fused with a polyhistidine tag at the C-terminus, constructed the plasmid 1; A DNA sequence encoding the human B2M (P61769) (Met1-Met119) constructed the plasmid 2. The two plasmids were co-expressed and the CD1B/B2M heterodimer was purified.
The recombinant heterodimer of human CD1B/B2M comprises 397 (297+100) amino acids and has a calculated molecular mass of 44.7 (33.0+ 11.7) KDa. The apparent molecular mass of rh CD1B/B2M heterodimer is approximately 39.6 and 13.1 KDa respectively in SDS-PAGE under reducing conditions.
Human CD1B & B2M Heterodimer HEK293 Overexpression Lysate: 使用指南
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.