This Human Nectin-2 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of Nectin-2 protein (Cat: 10005-H08H) from the overexpression lysate was verified.
A DNA sequence encoding the extracellular domain (Met 1-Leu 360) of human CD112 (NP_002847.1) was expressed with a C-terminal polyhistidine tag.
The recombinant mature human CD112 consists 340 amino acids and has a calculated molecular mass of 36.2 kDa. As a result of glycosylation, the recombinant protein migrates as an approximately 48 kDa protein in SDS-PAGE under reducing conditions.
Human Nectin-2 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.
Human Nectin-2 HEK293 Overexpression Lysate: 别称
Human CD112 Overexpression Lysate; Human HVEB Overexpression Lysate; Human Nectin-2 Overexpression Lysate; Human PRR2 Overexpression Lysate; Human PVRR2 Overexpression Lysate
Cluster of Differentiation 112 (CD112), also known as poliovirus receptor related protein 2 (PVRL2 or PRR2), is a single-pass type I transmembrane glycoprotein belonging to the Immunoglobulin superfamily. CD112 protein also serves as an entry for certain mutant strains of herpes simplex virus and pseudorabies virus, and thus is involved in cell to cell spreading of these viruses. CD112 protein has been identified as the ligand for DNAM-1 (CD226), and the interaction of CD226/CD112 protein can induce NK cell- and CD8+ T cell-mediated cytotoxicity and cytokine secretion. CD112 has been regarded as a critical component in allergic reactions, and accordingly may function as a novel target for anti-allergic therapy.
poliovirus receptor-related 2 (herpesvirus entry mediator B)
Bachelet I, et al. (2006) Mast cell costimulation by CD226/CD112 (DNAM-1/Nectin-2): a novel interface in the allergic process. J Biol Chem. 281(37): 27190-6.
Wang L, et al. (2009) Molecular cloning, characterization and three-dimensional modeling of porcine nectin-2/CD112. Vet Immunol Immunopathol. 132(2-4): 257-63.