|Product Description||SARS-CoV-2 Spike (Omicron BA.2.75) Fluc Pseudovirus|
|Envelope protein||SARS-CoV-2 Spike Protein (Omicron BA.2.75 | B.1.1.529)|
|Physical appearance||Light red to light yellow transparent liquid|
Pseudovirus Neutralization Assay
a. Mix 89% DMEM medium, 10% Fetal bovine serum and 1% Penicillin-Streptomycin to prepare complete DMEM medium.
b. Thaw the pseudovirus at room temperature.
c. Dilute your samples with complete DMEM medium in a 96-well white flat bottom plate to reach a volume of 50 ~ 140 μL per well, then add 10 ~ 50 μL pseudovirus suspension per well to reach a final volume of 100 ~ 150 μL per well. Gently flap to mix well. Incubate the plate in a 5% (vol/vol) CO2, 37°C incubator for 60 min.
d. Digest and resuspend HEK293/Human ACE2 Overexpression Stable Cells (ACROBiosystems, Cat. No. CHEK-ATP042) with complete DMEM medium. Adjust the cell density to 2 ~ 5 × 105 cells per milliliter with complete DMEM medium. Seed 100 μL the cell suspension per well into the 96-well plate. Gently flap to mix well. Incubate the plate in a 5% (vol/vol) CO2, 37°C incubator for 48 ~ 72 h. Notably, if a relative small cell number is used (e.g., 2 × 105 cells per milliliter), a relative long incubation time (e.g., 72 h) is reasonable; if a relative large cell number is used (e.g., 4 ~ 5 × 105 cells per milliliter), a relative short incubation time (e.g., 48 h) is reasonable.
e. Prepare the detection reagent (britelite plus Reporter Gene Assay System (PerkinElmer, Cat. No. 6066761)) and balance it to room temperature.
f. Take out the 96-well plate and discard 100 ~ 150 μL medium per well. Balance the plate to room temperature for 10 min. Add 100 μL detection reagent and mix well. Incubate for 2 min at room temperature.
g. Read the luminescence values (RLU) of the wells with a luminescence meter (PerkinElmer, Cat. No. HH34000000).
SARS-CoV-2 Spike (Omicron BA.2.75) Fluc Pseudovirus uses pseudotyped HIV-1 virus with firefly luciferase gene as the backbone and takes SARS-CoV-2 Spike protein as its envelope protein. It can effectively infect human ACE2 overexpressing cells and can be used in determining neutralizing antibody titer, screening for inhibitors of the Spike-ACE2 interaction, studying virus invasion and COVID-19 vaccine development.
Mutation(s) compared to wild type surface glycoprotein [severe acute respiratory syndrome coronavirus 2] (Accession # QHD43416.1): T19I, LPP24-26del, A27S, G142D, K147E, W152R, F157L, I210V, V213G, G257S, G339H, S371F, S373P, S375F, T376A, D405N, R408S, K417N, N440K,G446S, N40K, S477N, T478K, E484A, Q498R, N501Y, Y505H, D614G, H655Y, N679K, P681H, N764K, D796Y, Q954H, N969K ("LPP24-26del, A27S" equals to "L24S, PPA25-27del").
a. Though pseudovirus particles has no pathogenicity and cannot replicate, the assays should be carried out carefully in a Biosafety Level 2 or higher-level laboratory with a biosafety cabinet.
b. Serum samples from animals or humans should be inactivated in a water bath at 56°C for 30 min before being tested.
c. Please avoid freezing and thawing, which would influence the titer of the pseudovirus.
d. The product is for Research Use Only.
For more information and advice, please check the Data Sheet of this product.
Optional Relative Products
HEK293/Human ACE2 Stable Cell Line (ACROBiosystems, Cat. No. CHEK-ATP042)
The stable cell line is susceptible to SARS-CoV-2 Spike Fluc-GFP Pseudovirus.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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