This Mouse CD36 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD36 protein (Cat: 50422-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain of mouse CD36 (NP_001153030.1) (Gly 30-Lys 439) was expressed, with a polyhistidine tag at the C-terminus and a signal peptide at the N-terminus.
The secreted recombinant mouse CD36 consists of 421 amino acids and has a calculated molecular mass of 47.8 kDa. As a result of glycosylation, the apparent molecular mass of rmCD36 is approximately 80-90 kDa in SDS-PAGE under reducing conditions.
Mouse CD36 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.
The cluster of differentiation (CD) system is commonly used as cell markers in immunophynotyping. Different kinds of cells in the immune system can be identified through the surface CD molecules which associating with the immune function of the cell. There are more than 32 CD unique clusters and subclusters have been identified. Some of the CD molecules serve as receptors or ligands important to the cell through initiating a signal cascade which then alter the behavior of the cell. Some CD proteins do not take part in cell signal process but have other functions such as cell adhesion. Cluster of differentiation 36 (CD36), also known as FAT, SCARB3, GP88, glycoprotein IV (gpIV) and glycoprotein IIIb (gpIIIb), is a member of the CD system as well as the class B scavenger receptor family of cell surface proteins. CD36 can be found on the surface of many cell types in vertebrate animals and it consists of 472 amino acids and is extensively glycosylated. It is an integral membrane protein primarily serving as receptors for thrombospondin and collagen and by the erythrocytes infected with the human malaria parasite. The role of CD36 as a cell surface receptor has been extended to that of a signal transduction molecule.
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