Human NKp30/NCR3 HEK293 Overexpression Lysate

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Human NKp30/NCR3 HEK293 Overexpression Lysate: Product Information

Product Description
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-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the extracellular domain (Met 1-Gly 135) of human NCR3 (NP_667341.1) precursor was fused with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human NCR3 consists of 128 amino acids after removal of the signal peptide and has a predicted molecular mass of 14.4 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rh NCR3 is approximately 22-28 kDa due to glycosylation.

Human NKp30/NCR3 HEK293 Overexpression Lysate: Usage Guide

Preparation Method
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.
Lysis Buffer
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.
Recommend Usage
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.
Sample Buffer
1 X Sample Buffer (1 X modified RIPA buffer+1 X SDS loading buffer).
Stability & Storage
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.
Application
Western Blot (WB)
Optimal dilutions/concentrations should be determined by the end user.

Human NKp30/NCR3 HEK293 Overexpression Lysate: Alternative Names

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 Background Information

Natural Cytotoxicity Triggering Receptor 3, NCR3, also known as NKp3, 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 NKp3 has remained elusive, but the membrane-associated heparan sulfate (HS) proteoglycans are involved in the recognition of cellular targets by NKp3 was recently reported. NKp3 is a member of the immunoglobulin superfamily and one of three existing natural cytotoxicity-triggering receptors. NKp3 is a glycosylated protein and is thought to be selectively expressed in resting and activated natural killer cells. NKp3 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.
Full Name
natural cytotoxicity triggering receptor 3
References
  • 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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