This Mouse Secretogranin 3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Secretogranin 3 protein (Cat: 51561-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse Scg3 (NP_033156.1) (Met1-Leu471) was expressed with a polyhistidine tag at the C-terminus.
The recombinant mouse Scg3 consists of 460 amino acids and predicts a molecular mass of 52.4 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.
SCG3, also known as secretogranin 3, is a member of the chromogranin/secretogranin family. Members of this family may serve as precursors for biologically active peptides. SCG3 is transported to secretory granules (SGs) in neuroendocrine cells. SCG3 binds strongly to chromogranin A (CgA) in an intragranular milieu and targets CgA to SGs in pituitary and pancreatic endocrine cells. With a sucrose density gradient of rat insulinoma-derived INS-1 cell homogenates, SgIII is localized to the SG fraction and is fractionated to the SG membrane (SGM) despite lacking the transmembrane region.
Rong YP. et al., 2002, Sheng Wu Wu Li Xue Bao. 34 (4): 411-7.