This Mouse CD226 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD226 protein (Cat: 50232-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse DNAM1 (NP_848802.2) extracellular domain (Met 1-Pro 254) was fused with a polyhistidine tag at the C-terminus.
The recombinant mouse DNAM1 consists of 247 amino acids and has a predicted molecular mass of 28.2 kDa. As a result of glycosylation, the apparent molecular mass of rm DNAM1 is approximately 43 kDa in SDS-PAGE under reducing conditions.
Mouse CD226 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 Immunophenotyping. 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 320 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. CD226, also known as PTA1 or DNAM-1, is a member of the immunoglobulin superfamily containing 2 Ig-like domains of the V-set. High rate of CD226 (Cluster of Differentiation 226) is found on the surface of natural killer cells, platelets, monocytes and a subset of T cells. CD226 have binding sites with CD112 and CD155 and mediate cellular adhesion to other cells containing its ligands.