This Mouse CD5L overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD5L protein (Cat: 50020-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Val 352) of mouse CD5L (NP_033820.2) precursor was expressed with a polyhistidine tag at the C-terminus.
The secreted recombinant mouse CD5L consists of 342 amino acids and has a predicted molecular mass of 38 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rm CD5L is approximately 55-60 kDa due to glycosylation.
Mouse CD5L 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.
CD5L, also known as CD5 antigen-like, is a soluble protein belonging to group B of the scavenger receptor cysteine-rich (SRCR) superfamily and contains three SRCR domains. It is a secreted glycoprotein and expressed by macrophages presentin lymphoid tissues (spleen, lymph node, thymus, and bone marrow). It binds to myelomonocytic and lymphoid cells and may play an important role in the regulation of the innate and adaptive immune systems. CD5L functions as a pattern recognition molecule by binding both lipoteichoic acid (LTA) on Gram positive and lipopolysaccharide (LPS) on Gram negative bacteria. and the SRCR domain 1 of CD5L retains both the LPS and LTA binding activities. In addtion, it is revealed that CD5L seems to play a role as an inhibitor of apoptosis.
Resnick, D. et al., 1994, Trends Biochem. Sci. 19: 5-8.
Gebe, J. A. et al., 1997, J. Biol. Chem. 272 (10): 6151–6158.
Sarrias, M.R. et al., 2004, Crit. Rev. Immunol. 24: 1-37.
Mukhopadhyay, S. and Gordon, S., 2004, Immunobiology 209: 39-49.
Sarrias, M. R. et al.,2005, J. Biol. Chem. 280 (42): 35391–35398.