This Cynomolgus B7-H6 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of B7-H6 protein (Cat: 90802-C02H) from the overexpression lysate was verified.
A DNA sequence encoding the cynomolgus NCR3LG1 (XP_005578557.1) (Met1-Asp259) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant cynomolgus NCR3LG1 consists of 473 amino acids and predicts a molecular mass of 53.2 kDa.
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.