Mouse CD31 HEK293 Overexpression Lysate: Product Information
This Mouse CD31 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD31 protein (Cat: 50408-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse PECAM1 (NP_033868.3) extracellular domain (Met 1-Lys 590) was expressed, with a C-terminal polyhistidine tag.
The secreted recombinant mouse PECAM1 consists of 584 amino acids and has a calculated molecular mass of 66 kDa. The apparent molecular mass of the recombinant protein is approximately 85-95 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.
The Cluster of Differentiation 31 (CD31) adhesion molecule, also known as platelet-endothelial cell adhesion molecule-1 (PECAM-1), is the only known member of the CAM family on platelets. CD31 protein is a 13-kDa transmembrane glycoprotein expressed by endothelial cells, platelets, monocytes, neutrophils, and certain T cell subsets. CD31 protein is also expressed in certain tumors, including epithelioid hemangioendothelioma, other vascular tumors, and histiocytic malignancies. CD31 plays a key role in removing aged neutrophils and tissue regeneration. CD31 protein mediates the homotypic or heterotypic cell adhesion by binding to itself or the leukocyte integrin αvβ3, and thus plays a role in neutrophil recruitment in inflammatory responses, transendothelial migration of leukocytes, as well as in cardiovascular development.
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