This Mouse REG2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of REG2 protein (Cat: 51151-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse REG2 (NP_033069.1) (Met1-Ala173) was expressed with a C-terminal polyhistidine tag.
The recombinant mouse REG2 comprises 162 amino acids and has a predicted molecular mass of 18.3 kDa. The apparent molecular mass of the protein is approximately 19.1 kDa in SDS-PAGE under reducing conditions.
Mouse REG2 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.
Regenerating islet-derived proteins (Regs) comprise the superfamily of C-type lectin proteins encoded by Reg1, Reg2, Reg3a, Reg3b, Reg3g, Reg3d, and Reg4. The Reg family proteins are expressed in various tissues and have pleiotropic functions. Reg3b and Reg3g encode murine Reg3β and Reg3γ, respectively, and are murine homolog of human REG3A. Both proteins are highly expressed in the small intestine of adult mice at both mRNA and protein levels, but their expression is very low in the colon. Intriguingly, expression of Reg3β and Reg3γ is not detectable in the embryonic mouse intestine but gradually increases along with colonization of the commensal bacteria after birth.