Rat Nectin-2 HEK293 Overexpression Lysate: Product Information
This Rat 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: 80327-R08H) from the overexpression lysate was verified.
A DNA sequence encoding the rat PVRL2 (Q9ESS6) (Met1-Gly351) was expressed, fused with a polyhistidine tag at the C-terminus.
The recombinant rat PVRL2 comprises 331 amino acids and predicts a molecular mass of 36 kDa. The apparent molecular mass of the recombinant protein is approximately 46 kDa in SDS-PAGE under reducing conditions due to glycosylation.
Rat Nectin-2 HEK293 Overexpression Lysate: Usage Guide
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).
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.
Western Blot (WB) Optimal dilutions/concentrations should be determined by the end user.
Nectin-2 Background Information
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)
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