Mouse CD27 HEK293 Overexpression Lysate: Product Information
This Mouse CD27 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD27 protein (Cat: 50110-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Arg 182) of mouse CD27 (NP_001028298.1) was fused with a polyhistidine tag at the C-terminus.
The recombinant mouse CD27 consists of 173 amino acids after removal of the signal peptide and has a predicted molecular mass of 19.6 kDa. In SDS-PAGE under reducing conditions, rmCD27 migrates as an approximately 35 kDa band due to glycosylation.
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.
CD27, also known as TNFRSF7, is a member of the TNF-receptor superfamily limited to cells of the lymphoid lineage, and exists as both a dimeric glycoprotein on the cell surface and as a soluble protein in serum. As a type I transmembrane glycoprotein of about 55 kDa existing as disulfide-linked homodimer, CD27 has been shown to play roles in lymphoid proliferation, differentiation, and apoptosis.It has important role in generation of T cell immunity, and is an apparently robust marker for normal memory B cells. It is a T and B cell co-stimulatory molecule, the activity of CD27 is governed by its TNF-like ligand CD7 on lymphocytes and dendritic cells. The CD27-CD7 interaction is required for Th1 generation responses to differentiation signals and long-term maintenance of T cell immunity, and meanwhile, plays a key role in regulating B-cell differentiation, activation and immunoglobulin synthesis.
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