This Mouse APRIL/TNFSF13 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of APRIL/TNFSF13 protein (Cat: 50965-M01H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse TNFSF13 (Q9D777) (Ala96-Leu240) was expressed with the fused Fc region of human IgG1 at the N-terminus.
The recombinant mouse TNFSF13/Fc comprises 405 amino acids and has a predicted molecular mass of 44.8 kDa. The apparent molecular mass of the protein is approximately 47 kDa in SDS-PAGE under reducing conditions 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.
TNFSF13 is a member of the tumor necrosis factor (TNF) ligand family. It is a ligand for TNFRSF17/BCMA. TNFSF13 is lowly expressed in normal tissues, but is elevated in several types of tumors and transformed cell lines. It is important for B cell development. TNFSF13 may also play a role in T-independent type II antigen responses and T cell survival, and induce proliferation/survival of non lymphoid cells. It exists as a functional homotrimer. It can bind to two cell surface receptors, BCMA and TACI, which it shares with BAFF to exert downstream T- and B-cell regulatory effects. TNFSF13 also has been demonstrated to bind to proteoglycans on the cell surface.
tumor necrosis factor (ligand) superfamily, member 13
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