This Mouse LAG3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of LAG3 protein (Cat: 53069-M08H) from the overexpression lysate was verified.
A DNA sequence encoding the mouse Lag3 (NP_032505.1) (Met1-Leu442) was expressed with a polyhistidine tag at the C-terminus.
The recombinant mouse Lag3 consists 431 amino acids and predicts a molecular mass of 46.8 kDa.
Mouse LAG3 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.
LAG3 (Lymphocyte Activating 3) is a Protein Coding gene. 2 alternatively spliced human isoforms have been reported. LAG3, also known as CD223 and Lymphocyte activation gene 3, belongs to the immunoglobulin (Ig) superfamily. The LAG3 gene contains 8 exons. It is selectively expressed in activated T and NK cells. LAG3 contains 4 extracellular Ig-like domains and has a negative regulatory function in T cells. It also acts as a new marker of T cell-induced B cell activation. As a soluble molecule, LAG3 activates antigen-presenting cells through MHC class II signaling, leading to increased antigen-specific T-cell responses in vivo. Diseases associated with LAG3 include Smoldering Myeloma and Kyphoscoliotic Heart Disease.