This Human GITR overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of GITR protein (Cat: 13643-H02H) from the overexpression lysate was verified.
A DNA sequence encoding the human TNFRSF18 (Q9Y5U5-1) (Met1-Glu161) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant human TNFRSF18/Fc is a disulfide-linked homodimer. The reduced monomer comprises 377 amino acids and has a predicted molecular mass of 41.5 kDa. The apparent molecular mass of the protein is approximately 96 and 45 kDa in SDS-PAGE under reducing conditions.
Human GITR 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.
1. Centrifuge the tube for a few seconds and ensure the pellet at the bottom of the tube.
2. Re-dissolve the pellet using 200μL pure water and boil for 2-5 min.
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
稳定性 & 储存条件
Store at 4℃ for up to twelve months from date of receipt. After re-dissolution, aliquot and store at -80℃ for up to twelve months. Avoid repeated freeze-thaw cycles.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Human GITR HEK293 Overexpression Lysate: 别称
Human AITR Overexpression Lysate; Human CD357 Overexpression Lysate; Human GITR Overexpression Lysate; Human GITR-D Overexpression Lysate
GITR, also known as TNFRSF18(CD357), belongs to the tumor necrosis factor receptor (TNF-R) superfamily. It is the receptor for TNFSF18. GITR plays a key role in dominant immunological self-tolerance maintained by CD25(+)CD4(+) regulatory T cells. GITR may be involved in interactions between activated T-lymphocytes and endothelial cells and in the regulation of T-cell receptor-mediated cell death. GITR and its ligand are important costimulatory molecules in the pathogenesis of autoimmune diseases. It also mediates NF-kappa-B activation via the TRAF2/NIK pathway.