This Mouse CD10 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD10 protein (Cat: 50218-M07H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Tyr 52-Trp 750) of mouse Mme (NP_032630.2) was fused with a polyhistidine tag at the N-terminus.
The secreted recombinant mouse Mme consists of 715 amino acids and has a calculated molecular mass of 82.2 kDa. As a result of glycosylation, the recombinant protein migrates as an approximately 90-100 kDa protein in SDS-PAGE under reducing conditions.
Mouse CD10 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 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 alters the behavior of the cell. Some CD proteins do not take part in the cell signal process but have other functions such as cell adhesion. The cluster of differentiation 10 (CD10), also known as Neprilysin and neutral endopeptidase, is a member of the CD system. CD10 is a zinc-dependent metalloprotease enzyme that had the function to degrade some small secreted peptides such as the amyloid beta-peptide. It exists as a membrane-bound protein and has a high concentration in kidney and lung tissues. Mutations in the CD10 gene can induce the familial forms of Alzheimer's disease, providing strong evidence for the protein's association with the Alzheimer's disease process. CD10 is also associated with other biochemical processes.
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