Rat NKp46/NCR1 HEK293 Overexpression Lysate

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Rat NKp46/NCR1 HEK293 Overexpression Lysate: 产品信息

产品描述
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.
表达宿主
HEK293 Cells
种属
Rat
蛋白构建信息
A DNA sequence encoding the rat NCR1 (Q9Z0H5) (Met1-Asn255) was expressed, fused with the Fc region of human IgG1 at the C-terminus.
分子量
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

制备方法
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.
裂解缓冲液
Modified RIPA Lysis Buffer: 50 mM Tris-HCl pH 7.4, 150 mM NaCl, 1mM EDTA, 1% Triton X-100, 0.1% SDS, 1% Sodium deoxycholate, 1mM PMSF.
使用建议
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.
缓冲液
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.
应用
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

NKp46/NCR1 背景信息

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.
全称
natural cytotoxicity triggering receptor 1
参考文献
  • Carbone E, et al. (2005) HLA class I, NKG2D, and natural cytotoxicity receptors regulate multiple myeloma cell recognition by natural killer cells. Blood. 105(1):251-8.
  • Sivori S, et al. (1997) p46, a Novel Natural Killer Cell-specific Surface Molecule That Mediates Cell Activation. J Exp Med. 186(7):1129-36.
  • Biassoni R, et al. (2004) Human natural killer cell receptors: insights into their molecular function and structure. J Cell Mol Med. 7(4):376-87.
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