This Rhesus CD160 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of CD160 protein (Cat: 90224-C02H) from the overexpression lysate was verified.
A DNA sequence encoding the rhesus CD160 (XP_001089019.1) (Met1-Leu158) was expressed with the Fc region of human IgG1 at the C-terminus.
The recombinant Rhesus CD160 is a disulfide-linked homodimer. The reduced monomer comprises 375 amino acids and has a calculated molecular mass of 41.8 KDa.The apparent molecular mass of the protein is approximately 47-52 KDa in SDS-PAGE.
Rhesus CD160 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.
CD16 antigen, also known as Natural killer cell receptor BY55 and CD16, is a cell membrane protein which contains one Ig-like V-type (immunoglobulin-like) domain. CD16 is a GPI-anchored lymphocyte surface receptor in which expression is mostly restricted to the highly cytotoxic CD56(dim)CD16(+) peripheral blood NK subset. CD16 is a receptor showing broad specificity for both classical and non-classical MHC class I molecules. CD16 is expressed in spleen, peripheral blood, and small intestine. Expression of CD16 is restricted to functional NK and T cytotoxic lymphocytes. CD16 acts as a co-activator receptor for CD3-induced proliferation of CD4+ CD16+ T cells isolated from inflammatory skin lesions. Unique CD4+ CD16+ lymphocyte subset may play a role in the pathogenesis of skin inflammation. Activated NK lymphocytes release a soluble form of CD16 that functionally impairs the MHC-I-specific cytotoxic CD8(+) T lymphocyte responsiveness.