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Testing Data
(Figure 1. Illustration of the replication-deficient MLV particle pseudotyped with SARS-CoV-2 Spike protein)
COVID 19 Spike 614G Coronavirus Pseudovirus | COVID-19 pseudovirus
SARS-CoV-2 Pseudoviral Particles, 614G
Synonyms
COVID 19 Spike 614G Coronavirus; SARS-CoV-2 Pseudoviral Particles; 614G; 2019 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 614G; SARS-CoV-2 Pseudoviral Particles Spike 614G; COVID-19 pseudovirus
Application Notes
Our Pseudovirus Particles generate robust chemiluminescent signals in cell assays when coupled with our firefly luciferase assay kit (Catalog# MBS434279), useful for 1) screening potential inhibitor to block SARS-CoV-2 entry and viral protein translation; 2) measuring the activity of and screening for neutralizing antibody against SARS-CoV-2-614G.
ASSAY PROTOCOL I (Fig. 2 & 3)
Note: requires a luciferase assay reagent. (Catalog# MBS434279)
Cell Infection:
1. Count HEK293-ACE2 cells (Catalog# MBS434276) to be infected and seed ~20K cells per well into 96-well plates (50 ul per well) DMEM with 10% FC II Serum (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 50 ul SARS-CoV-2 pseudoviral particles into each well (12.5 ul 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 cells
1. 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.
ASSAY PROTOCOL II (Fig. 4)
Note: requires a luciferase assay reagent. (Catalog# MBS434279)
Cell Infection:
1. Count HEK293-ACE2 cells (Catalog # MBS434276) to be infected and seed ~7.5K cells per well into 384-well plates (15 ul per well) in DMEM with 10% FC II Serum (no antibiotics).
2. On the 2nd day, dilute the antibodies (5X of the final concentrations) in DMEM with 10% FC II Serum (no antibiotics).
Note: for most antibodies, we recommend testing 11 different doses (1:3 serial dilutions), starting from 30 ug/ml or 10 ug/ml (final concentration in the well).
3. Take 5 ul of the antibody solution and add into a compound plate containing 12.5 ul of SARS-CoV-2-PP per well (i.e. SARS-CoV-2-614G PP, Catalog# MBS434278).
4. Incubate at 37°C for 1 hr.
5. Take a cell plate out from the incubator, remove the medium, add 17.5 ul of SARS-CoV-2-PP and antibody mix into each well of the cell plate. Centrifuge at 700 rpm for 15 min at 4°C.
6. Add additional 7.5 ul of DMEM with 10% FC into each well.
7. Incubate at 37°C for 42 hr.
Measurement of Luciferase Activity in Infected cells
1. Remove supernatant.
2. Add 20 ul luciferase assay reagent into each well.
3. Read in a luminescence plate reader and record the data.
ASSAY PROTOCOL I (Fig. 2 & 3)
Note: requires a luciferase assay reagent. (Catalog# MBS434279)
Cell Infection:
1. Count HEK293-ACE2 cells (Catalog# MBS434276) to be infected and seed ~20K cells per well into 96-well plates (50 ul per well) DMEM with 10% FC II Serum (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 50 ul SARS-CoV-2 pseudoviral particles into each well (12.5 ul 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 cells
1. 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.
ASSAY PROTOCOL II (Fig. 4)
Note: requires a luciferase assay reagent. (Catalog# MBS434279)
Cell Infection:
1. Count HEK293-ACE2 cells (Catalog # MBS434276) to be infected and seed ~7.5K cells per well into 384-well plates (15 ul per well) in DMEM with 10% FC II Serum (no antibiotics).
2. On the 2nd day, dilute the antibodies (5X of the final concentrations) in DMEM with 10% FC II Serum (no antibiotics).
Note: for most antibodies, we recommend testing 11 different doses (1:3 serial dilutions), starting from 30 ug/ml or 10 ug/ml (final concentration in the well).
3. Take 5 ul of the antibody solution and add into a compound plate containing 12.5 ul of SARS-CoV-2-PP per well (i.e. SARS-CoV-2-614G PP, Catalog# MBS434278).
4. Incubate at 37°C for 1 hr.
5. Take a cell plate out from the incubator, remove the medium, add 17.5 ul of SARS-CoV-2-PP and antibody mix into each well of the cell plate. Centrifuge at 700 rpm for 15 min at 4°C.
6. Add additional 7.5 ul of DMEM with 10% FC into each well.
7. Incubate at 37°C for 42 hr.
Measurement of Luciferase Activity in Infected cells
1. Remove supernatant.
2. Add 20 ul luciferase assay reagent into each well.
3. Read in a luminescence plate reader and record the data.
Features
• Robust: Excellent signal to noise (basal) ratio
• Easy to use: Amenable to HTS format (96-well, 384-well and 1536-well format)
• Easy to use: Amenable to HTS format (96-well, 384-well and 1536-well format)
Contents
10 ml (2 tubes, 5 mL/tube), for 2 multi-well plates
Shipping Note
MBS434278 is available for shipment in the United States, Canada and European countries. Please inquire for shipment to other countries.
Dry Ice Shipment
Extra charge fee may add to your shipping cost as dry ice is required to ship this product.
Preparation and Storage
Upon receiving this item , store at -70°C right away. Thaw each vial immediately before each use (don't aliquot and refreeze)
Shelf life : Six months from the date of shipping when stored at -70°C.
Shelf life : Six months from the date of shipping when stored at -70°C.
Testing Data
(Figure 1. Illustration of the replication-deficient MLV particle pseudotyped with SARS-CoV-2 Spike protein)
Testing Data
(Figure 1. Illustration of the replication-deficient MLV particle pseudotyped with SARS-CoV-2 Spike protein)
Testing Data
(Figure 2. Pseudoviral Particle (PP) Infection Assays. SARS2-CoV-2-614D-pp, the PP pseudotyped with the 614D (original) spike protein (Catalog# MBS434275); SARS2-CoV-2-614G-pp, the PP pseudotyped with the 614G spike protein (Catalog# MBS434278).)
Testing Data
(Figure 2. Pseudoviral Particle (PP) Infection Assays. SARS2-CoV-2-614D-pp, the PP pseudotyped with the 614D (original) spike protein (Catalog# MBS434275); SARS2-CoV-2-614G-pp, the PP pseudotyped with the 614G spike protein (Catalog# MBS434278).)
Testing Data
(Figure 3. SARS-CoV-2 Viral Infection Inhibiting Test by Neutralization Antibodies. HEK293-ACE2 cells infected with SARS-CoV-2 Pseudoviral Particles (Catalog# MBS434278) under various amount of neutralizing antibodies (MBS434285).)
Testing Data
(Figure 3. SARS-CoV-2 Viral Infection Inhibiting Test by Neutralization Antibodies. HEK293-ACE2 cells infected with SARS-CoV-2 Pseudoviral Particles (Catalog# MBS434278) under various amount of neutralizing antibodies (MBS434285).)
Testing Data
(Figure 4. Antibody (Ab) Neutralization Assays with SARS-CoV-2-614G PP)
Testing Data
(Figure 4. Antibody (Ab) Neutralization Assays with SARS-CoV-2-614G PP)
Related Product Information for COVID-19 pseudovirus
It has been known that SARS coronavirus 2 (SARS-CoV-2) uses 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 receptor human ACE2 (hACE2) 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. They also contain the ORF for firefly luciferase as a reporter. They establish a pseudovirus entry assay for SARS-CoV-2 as the S 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 new SARS-CoV-2 strain with an amino acid change at position 614 from Asp to Gly in the viral S protein predominated over time in locales where it was found. It is reported by the Choe lab in The Scripps Research Institute that this currently dominate new strain with the D614G variation in the S protein “reduces S1 shedding and increases infectivity”. To support the study of this new SARS-CoV-2 strain, we established the SARS-CoV-2-614G Pseudoviral Particles with the spike protein carrying the 614G genotype (Genbank Accession #QHD43416.1, p.614D->G)
Our SARS-CoV-2 Pseudoviral Particles are replication-deficient MLV pseudotyped with the SARS-CoV-2 spike protein. They also contain the ORF for firefly luciferase as a reporter. They establish a pseudovirus entry assay for SARS-CoV-2 as the S 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 new SARS-CoV-2 strain with an amino acid change at position 614 from Asp to Gly in the viral S protein predominated over time in locales where it was found. It is reported by the Choe lab in The Scripps Research Institute that this currently dominate new strain with the D614G variation in the S protein “reduces S1 shedding and increases infectivity”. To support the study of this new SARS-CoV-2 strain, we established the SARS-CoV-2-614G Pseudoviral Particles with the spike protein carrying the 614G genotype (Genbank Accession #QHD43416.1, p.614D->G)
NCBI and Uniprot Product Information
Similar Products
Product Notes
The COVID-19 (Catalog #AAA434278) is a Pseudovirus and is intended for research purposes only. The product is available for immediate purchase. Our Pseudovirus Particles generate robust chemiluminescent signals in cell assays when coupled with our firefly luciferase assay kit (Catalog# MBS434279), useful for 1) screening potential inhibitor to block SARS-CoV-2 entry and viral protein translation; 2) measuring the activity of and screening for neutralizing antibody against SARS-CoV-2-614G. ASSAY PROTOCOL I (Fig. 2 & 3) Note: requires a luciferase assay reagent. (Catalog# MBS434279) Cell Infection: 1. Count HEK293-ACE2 cells (Catalog# MBS434276) to be infected and seed ~20K cells per well into 96-well plates (50 ul per well) DMEM with 10% FC II Serum (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 50 ul SARS-CoV-2 pseudoviral particles into each well (12.5 ul 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 cells 1. 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. ASSAY PROTOCOL II (Fig. 4) Note: requires a luciferase assay reagent. (Catalog# MBS434279) Cell Infection: 1. Count HEK293-ACE2 cells (Catalog # MBS434276) to be infected and seed ~7.5K cells per well into 384-well plates (15 ul per well) in DMEM with 10% FC II Serum (no antibiotics). 2. On the 2nd day, dilute the antibodies (5X of the final concentrations) in DMEM with 10% FC II Serum (no antibiotics). Note: for most antibodies, we recommend testing 11 different doses (1:3 serial dilutions), starting from 30 ug/ml or 10 ug/ml (final concentration in the well). 3. Take 5 ul of the antibody solution and add into a compound plate containing 12.5 ul of SARS-CoV-2-PP per well (i.e. SARS-CoV-2-614G PP, Catalog# MBS434278). 4. Incubate at 37°C for 1 hr. 5. Take a cell plate out from the incubator, remove the medium, add 17.5 ul of SARS-CoV-2-PP and antibody mix into each well of the cell plate. Centrifuge at 700 rpm for 15 min at 4°C. 6. Add additional 7.5 ul of DMEM with 10% FC into each well. 7. Incubate at 37°C for 42 hr. Measurement of Luciferase Activity in Infected cells 1. Remove supernatant. 2. Add 20 ul luciferase assay reagent into each well. 3. Read in a luminescence plate reader and record the data. Researchers should empirically determine the suitability of the COVID-19 for an application not listed in the data sheet. Researchers commonly develop new applications and it is an integral, important part of the investigative research process. It is sometimes possible for the material contained within the vial of "COVID 19 Spike 614G Coronavirus, Pseudovirus" to become dispersed throughout the inside of the vial, particularly around the seal of said vial, during shipment and storage. We always suggest centrifuging these vials to consolidate all of the liquid away from the lid and to the bottom of the vial prior to opening. Please be advised that certain products may require dry ice for shipping and that, if this is the case, an additional dry ice fee may also be required.Precautions
All products in the AAA Biotech catalog are strictly for research-use only, and are absolutely not suitable for use in any sort of medical, therapeutic, prophylactic, in-vivo, or diagnostic capacity. By purchasing a product from AAA Biotech, you are explicitly certifying that said products will be properly tested and used in line with industry standard. AAA Biotech and its authorized distribution partners reserve the right to refuse to fulfill any order if we have any indication that a purchaser may be intending to use a product outside of our accepted criteria.Disclaimer
Though we do strive to guarantee the information represented in this datasheet, AAA Biotech cannot be held responsible for any oversights or imprecisions. AAA Biotech reserves the right to adjust any aspect of this datasheet at any time and without notice. It is the responsibility of the customer to inform AAA Biotech of any product performance issues observed or experienced within 30 days of receipt of said product. To see additional details on this or any of our other policies, please see our Terms & Conditions page.Item has been added to Shopping Cart
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