This Rat NKp46/NCR1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NKp46/NCR1 protein (Cat: 80378-R02H) from the overexpression lysate was verified.
A DNA sequence encoding the rat NCR1 (Q9Z0H5) (Met1-Asn255) was expressed, fused with the Fc region of human IgG1 at the C-terminus.
The recombinant rat NCR1/Fc is a disulfide-linked homodimer. The reduced monomer comprises 480 amino acids and has a predicted molecular mass of 54.1 kDa. The apparent molecular mass of the protein is approximately 65 and 36 kDa in SDS-PAGE under reducing conditions.
Rat NKp46/NCR1 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.
Rat NKp46/NCR1 HEK293 Overexpression Lysate: 别称
Rat Ar1 Overexpression Lysate; Rat Kilr1 Overexpression Lysate; Rat Ly94 Overexpression Lysate; Rat NCR1 Overexpression Lysate
NCR1, also known as NK-p46 and CD335, is a natural cytotoxicity receptor(NCR). NCRs are type I transmembrane proteins with 1-2 extracellular immunoglobulin domains, a transmembrane domain containing a positively charged amino acid residue, and a short cytoplasmic tail. All are expressed almost exclusively by NK cells and play a major role in triggering NK-mediated killing of most tumor cell lines. NKp46 has two extracellular Ig-like domains followed by a ~40 residue stalk region, a type I transmembrane domain, and a short cytoplasmic tail. NKp46 has been implicated in NK cell-mediated lysis of several autologous tumor cells, pathogen-infected cell lines, and mononuclear phagocytes infected with an intracellular bacterium.