| Product Description | SARS-CoV-2 Spike (XBB.1.5) Fluc-GFP Pseudovirus |
| Backbone | HIV-1 |
| Envelope protein | SARS-CoV-2 Spike Protein (XBB.1.5) |
| Reporter | Firefly luciferase-GFP |
| Physical appearance | Dark red-brown transparent liquid |
| Storage | -70℃ |
| Transport | Dry ice |
| TableApplication | Neutralization assay |
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. Dilute the pseudovirus with complete DMEM medium according to your pre-test results. In general, we recommend 125-fold dilution if you use the same materials and luminescence meter as those in this protocol (e. g., 20 μL Pseudovirus + 2.48 mL complete DMEM medium). c. Dilute your samples with complete DMEM medium in a 96-well white flat bottom plate to reach a volume of 75 μL per well, then add 25 μL pseudovirus suspension per well to reach a final volume of 100 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 4 ~ 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 h. 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 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).
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. Dilute the pseudovirus with complete DMEM medium according to your pre-test results. In general, we recommend 125-fold dilution if you use the same materials and luminescence meter as those in this protocol (e. g., 20 μL Pseudovirus + 2.48 mL complete DMEM medium). c. Dilute your samples with complete DMEM medium in a 96-well white flat bottom plate to reach a volume of 75 μL per well, then add 25 μL pseudovirus suspension per well to reach a final volume of 100 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 4 ~ 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 h. 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 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).
Basic advice
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.
Please contact us via TechSupport@acrobiosystems.com if you have any question on this product.
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