This Mouse CD200 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD200 protein (Cat: 50074-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse CD200 (O54901.1) (Met 1-Gly 232) was expressed with a polyhistidine tag at the C-terminus.
The secreted recombinant mouse CD200 consists of 213 amino acids and has a calculated molecular mass of 24 kDa. As a result of glycosylation, the apparent molecular mass of rmCD38 is approximately 38-43 kDa in SDS-PAGE under reducing conditions.
Mouse CD200 HEK293 Overexpression Lysate: 用法
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.
CD2 (OX-2) is a cell surface glycoprotein that imparts immune privileges by suppressing alloimmune and autoimmune responses through its receptor, CD2R, expressed primarily on myeloid cells. Signals delivered through the CD2:CD2R axis have been shown to play an important role in the regulation of anti-tumor immunity, and overexpression of CD2 has been reported in a number of malignancies, including CLL, as well as on cancer stem cells. The role of CD2-CD2R signaling in immune regulation of the central nervous system has become a popular field of research in recent years. Many studies have shown that there is a close correlation between CD2-CD2R, microglia activation, and Parkinson's disease (PD). The ability of CD2 to suppress myeloid cell activation is critical for maintaining normal tissue homeostasis but may also enhance the survival of migratory neoplastic cells. CD2 and CD2R associate via their respective N-terminal Ig-like domains. CD2 has been characterized as an important immunoregulatory molecule, increased expression of which can lead to decreased transplant rejection, autoimmunity, and allergic disease. Elevated CD2 expression has been reported to be associated with poor prognosis in a number of human malignancies. In addition, CD2 also plays an important role in prevention of graft rejection, autoimmune diseases and spontaneous abortion.