This Mouse Oncostatin M/OSM overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Oncostatin M/OSM protein (Cat: 50112-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mature form of mouse OSM (P53347) (Met 1-Arg 206) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse OSM comprises 193 amino acids and has a calculated molecular mass of 22 kDa. As a result of glycosylation, the apparent molecular mass of the recombinant protein is approximately 35-40 kDa in SDS-PAGE under reducing conditions.
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.
Oncostatin M (OSM) is a glycoprotein belonging to the interleukin-6 family of cytokines that has functions mainly in cell growth. Oncostatin M (OSM) is considered as a pleiotropic cytokine that signals through cell surface receptors typeⅠand typeⅡ both of which share the similarity of containing protein gp130 and takes part in many bio metabolism processes including liver development, hematopoiesis, inflammation, bone formation, and destruction and possibly CNS development. Oncostatin M (OSM) was previously identified by its ability to inhibit the growth of cells from melanoma and other solid tumors. It also has been reported that OSM, like LIF, IL-6, and G-CSF, can inhibit the proliferation of murine M1 myeloid leukemic cells and can induce their differentiation into macrophage-like cells. The human form of OSM is insensitive between pH2 and 11 and resistant to heating for one hour at 56 degrees but is not stable at 90 degrees. The human OSM is produced as a precursor containing 252 amino acids, whose first 25 amino acids function as a secretory signal peptide and which on removal yields the soluble 227 amino acid pro-OSM. Removal of the C-terminal most 31 amino acids produces the fully active 196 residue form.
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