COVID-19 pseudovirus :: COVID 19 Spike Coronavirus Pseudovirus假病毒顆粒

Product NameCOVID 19 Spike Coronavirus (COVID-19), Pseudovirus Popular ItemFull Product NameSARS-CoV-2 Pseudoviral Particles SpikeProduct Synonym Names2019 Novel Coronavirus; Coronavirus; CoV; COVID-19 virus; HCoV-2; Human Coronavirus 2019; SARS2; SARS-CoV-2; Severe acute respiratory syndrome coronavirus 2; VLP; Virus-like particle; Spike; SARS-CoV-2 Pseudoviral Particles SpikeProduct Gene NameCOVID-19 pseudovirus[Similar Products]Product Synonym Gene Name2019-nCoV; SARS-CoV-2; SARS coronavirus 2 proteome; COVID19; SARS-CoV2
Features? Robust: Excellent signal to noise (basal) ratio? Easy to use: Amenable to HTS format (96-well, 384-well and 1536-well format)Contents10 ml (2 tubes, 5 mL/tube), for 2 multi-well plates
It has been known that the coronaviruses SARS-CoV-2 and SARS-CoV use human ACE2 as the entry receptor and human proteases as the entry activators. The virus surface spike (S) protein mediates SARS-CoV-2 entry into cells. To fulfill its function, SARS-CoV-2 spike binds to the human ACE2 (hACE2) receptor through its receptor-binding domain (RBD) and is proteolytically activated by human proteases.Our SARS-CoV-2 Pseudoviral Particles are replication-deficient MLV pseudotyped with the SARS-CoV-2 spike protein carrying the original D614 genotype (Genbank Accession #QHD43416.1). They also contain the ORF for firefly luciferase as a reporter. They establish a pseudovirus entry assay for SARS-CoV-2 as the spike protein mediated cell entry can be conveniently measured via the luciferase reporter activity. This pseudovirus assay isolates the SARS-CoV-2 viral entry from other steps of the viral infection cycle.A related item, Catalog MBS434278, is the SARS-CoV-2 virus pseudotyped with the 614G variant spike protein.Product Categories/Family for COVID-19 pseudovirusPseudovirusApplication Notes for COVID-19 pseudovirusCoupled with firefly luciferase assay kit the Pseudovirus infection generates robust chemiluminescent signals,1) for screening potential inhibitor to block SARS-CoV-2 entry and viral protein translation;2) for measuring the activity of and screening for neutralizing antibody against SARS-CoV-2.Assay ProtocolNote: requires a luciferase assay reagent (any commercially available or use Cat# MBS434279)Cell Infection:1. Count Vero E6 cells or HEK293-ACE2 cells (MBS434276) to be infected and seed ~20K cells per well into 96-well plates (50 ul per well) DMEM with 10% FC (no antibiotics) or 5K cells per well into 384-well plates (15 ul per well).2. Culture cells overnight to make sure the cells stably adhere to the plates.3. On the 2nd day, remove media, add 50ul SARS-CoV-2 Pseudoviral Particles into each well (12.5ul for 384-well plate). Spin at 700 rpm for 15 min at 4°C.Note: thaw the pseudoviral particles immediately before use (take about 1 hour to thaw in ice) and use it within 2 hours.4. Incubate for 2 hrs at 37°C.5. Add 50 ul DMEM with 10% FC into each well (12.5 ul for 384-well plates).6. Incubate for 48 hrs at 37°C.Measurement of Luciferase Activity in Infected cells1. Remove supernatant.2. Add 100 ul Luciferase assay reagent (20 ul for 384-well plates).3. Read in a luminescence plate reader, and record the data. (Note: the RLU values are higher from 96-well.)

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