Human NKp30/NCR3 HEK293 Overexpression Lysate

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Human NKp30/NCR3 HEK293 Overexpression Lysate: 产品信息

产品描述
This Human NKp30/NCR3 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of NKp30/NCR3 protein (Cat: 10480-H03H) from the overexpression lysate was verified.
表达宿主
HEK293 Cells
种属
Human
蛋白构建信息
A DNA sequence encoding the extracellular domain (Met 1-Gly 135) of human NCR3 (NP_667341.1) precursor was fused with the C-terminal polyhistidine-tagged Fc region of human IgG1 at the C-terminus.
分子量
The recombinant human NCR3/Fc is a disulfide-linked homodimer after removal of the signal peptide. The reduced monomer consists of 365 amino acids and has a predicted molecular mass of 41 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh NCR3/Fc monomer is approximately 50-55 kDa due to glycosylation.

Human NKp30/NCR3 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.

Human NKp30/NCR3 HEK293 Overexpression Lysate: 别称

Human 1C7 Overexpression Lysate; Human CD337 Overexpression Lysate; Human DAAP-90L16.3 Overexpression Lysate; Human LY117 Overexpression Lysate; Human MALS Overexpression Lysate; Human NCR3 Overexpression Lysate; Human NKp30 Overexpression Lysate

NKp30/NCR3 背景信息

Natural Cytotoxicity Triggering Receptor 3, NCR3, also known as NKp30, or CD337, is a natural cytotoxicity receptor, expressed on subsets of human peripheral blood NK cells, involved in NK cell killing of tumor cells and immature dendritic cells. The cellular ligand for NKp30 has remained elusive, but the membrane-associated heparan sulfate (HS) proteoglycans are involved in the recognition of cellular targets by NKp30 was recently reported. NKp30 is a member of the immunoglobulin superfamily and one of three existing natural cytotoxicity-triggering receptors. NKp30 is a glycosylated protein and is thought to be selectively expressed in resting and activated natural killer cells. NKp30 is a stimulatory receptor on human NK cells implicated in tumor immunity and is capable of promoting or terminating dendritic cell maturation. NCR3 may play a role in inflammatory and infectious diseases.
全称
natural cytotoxicity triggering receptor 3
参考文献
  • Warren HS, et al. (2005) Evidence that the cellular ligand for the human NK cell activation receptor NKp30 is not a heparan sulfate glycosaminoglycan. J Immunol. 175(1): 207-12.
  • Mulcahy H, et al. (2006) LST1 and NCR3 expression in autoimmune inflammation and in response to IFN-gamma, LPS and microbial infection. Immunogenetics. 57(12): 893-903.
  • Hsieh CL, et al. (2006) NKp30 is a functional activation receptor on a subset of rat natural killer cells. Eur J Immunol. 36(8): 2170-80.
  • Ponnampalam AP, et al. (2008) Identification and hormonal regulation of a novel form of NKp30 in human endometrial epithelium. Eur J Immunol. 38(1): 216-26.
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