Human PVRL1/NECTIN1 HEK293 Overexpression Lysate

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Human PVRL1/NECTIN1 HEK293 Overexpression Lysate: Product Information

Product Description
This Human PVRL1/NECTIN1 overexpression lysate was created in HEK293 Cells and intented for use as a Western blot (WB) positive control. Purification of PVRL1/NECTIN1 protein (Cat: 11611-H08H) from the overexpression lysate was verified.
Expression Host
HEK293 Cells
Species
Human
Sequence Information
A DNA sequence encoding the human PVRL1 isoform 1 (NP_002846.3) extracellular domain (Met 1-Thr 334) was expressed, with a polyhistidine tag at the C-terminus.
Molecule Mass
The recombinant human PVRL1 consists of 315 amino acids and predictes a molecular mass of 35.4 kDa. In SDS-PAGE under reducing conditions, the apparent molecular mass of rhPVRL1 is approximately 45-50 kDa due to glycosylation.

Human PVRL1/NECTIN1 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 PVRL1/NECTIN1 HEK293 Overexpression Lysate: Alternative Names

Human CD111 Overexpression Lysate; Human CLPED1 Overexpression Lysate; Human ED4 Overexpression Lysate; Human HIgR Overexpression Lysate; Human HV1S Overexpression Lysate; Human HVEC Overexpression Lysate; Human nectin-1 Overexpression Lysate; Human OFC7 Overexpression Lysate; Human PRR Overexpression Lysate; Human PRR1 Overexpression Lysate; Human PVRR Overexpression Lysate; Human PVRR1 Overexpression Lysate; Human SK-12 Overexpression Lysate

PVRL1/NECTIN1 Background Information

Poliovirus receptor-related 1 (herpesvirus entry mediator C; nectin-1; CD111), also known as PVRL1 is a cell adhesion molecule belonging to the immunoglobulin superfamily that can bind to virion glycoprotein D (gD) to mediate entry of herpes simplex viruses (HSV) and pseudorabies virus (PRV). CD111/Nectin-1/PVRL1 colocalizes with E-cadherin at adherens junctions in epithelial cells. The disruption of cell junctions can result in the redistribution of nectin-1. To determine whether disruption of junctions by calcium depletion influenced the susceptibility of epithelial cells to viral entry, Madin-Darby canine kidney cells expressing endogenous nectin-1 or transfected human nectin-1 were tested for the ability to bind soluble forms of viral gD and to be infected by HSV and PRV, before and after calcium depletion. It has been revealed that binding of HSV and PRV gD was localized to adherens junctions in cells maintained in normal medium but was distributed, along with nectin-1, over the entire cell surface after calcium depletion. Both the binding of gD and the fraction of cells that could be infected by HSV-1 and PRV were enhanced by calcium depletion. Taken together, CD111/Nectin-1/PVRL1 confined to adherens junctions in epithelial cells is not very accessible to virus, whereas dissociation of cell junctions releases nectin-1 to serve more efficiently as an entry recptor.
Full Name
poliovirus receptor-related 1 (herpesvirus entry mediator C)
References
  • Yoon M, et al. (2002) Disruption of adherens junctions liberates nectin-1 to serve as receptor for herpes simplex virus and pseudorabies virus entry. J Virol. 76(14): 7203-8.
  • Mata M, et al. (2001) HveC (nectin-1) is expressed at high levels in sensory neurons, but not in motor neurons, of the rat peripheral nervous system. J Neurovirol. 7(5): 476-80.
  • Haarr L, et al. (2001) Transcription from the gene encoding the herpesvirus entry receptor nectin-1 (HveC) in nervous tissue of adult mouse. Virology. 287(2): 301-9.
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