This Human CD31 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD31 protein (Cat: 10148-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Lys 601) of human CD31 (EAW94208.1) was expressed with a polyhistidine tag at the C-terminus.
The secreted recombinant human CD31 consists of 585 amino acids after removal and predicts a molecular mass of 66 kDa. As a result of glycosylation, the rhCD31 migrates as approximately 100-110 kDa band in SDS-PAGE under reducing conditions.
Human CD31 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.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human CD31 HEK293 Overexpression Lysate: 别称
Human CD31 Overexpression Lysate; Human CD31/EndoCAM Overexpression Lysate; Human endoCAM Overexpression Lysate; Human GPIIA' Overexpression Lysate; Human PECA1 Overexpression Lysate; Human PECAM-1 Overexpression Lysate
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 130-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.