A SARS-CoV-2 spike pseudotyped lentivirus kit was obtained through BEI Resources, NIAID, NIH (SARS-Related Coronavirus 2, Wuhan-Hu-1 Spike-Pseudotyped Lentiviral Kit V2, NR-53816). We used HEK-293T–expressing human angiotensin-converting enzyme 2, also known as HEK-293T–hACE2, which are susceptible to SARS-CoV-2. This cell line was obtained through BEI Resources, NIAID, NIH, NR-52511. Serial dilutions of sera were incubated with the SARS-CoV-2 spike pseudotyped lentivirus, following a protocol by Balazs and Bloom (50 ). Cells were lysed using luciferase cell culture lysis buffer (Promega). Luciferase reaction was performed using 30 μl of cell lysis (Promega). The reaction was added to 96-well black optiplates (PerkinElmer). Luminescence was measured using a PerkinElmer Victor3 luminometer.
96 well black optiplates
Manufactured by PerkinElmer
The 96-well black optiplates are a type of multi-well microplate designed for use in various laboratory applications. These plates have 96 individual wells arranged in a 8x12 grid configuration and are constructed with a black-colored material that minimizes light interference. The black color helps to reduce background fluorescence and luminescence, improving the signal-to-noise ratio for sensitive detection methods.
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Lab products found in correlation
4 protocols using 96 well black optiplates
1
SARS-CoV-2 Spike Pseudotyped Lentivirus Assay
2
SIV Neutralization Assay with TZM-bl Cells
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TZM-bl cells (a HeLa cell line engineered to express human receptors and coreceptors for HIV, in addition to a Tat-inducible luciferase gene) were used to measure vaccine-induced antibody neutralization of SIV. TZM-bl cells were cultured in T75 flasks (Thermo Scientific) at 104 cell/ml density for 3 d in 10% FBS complete DMEM (GIBCO). On the day of the assay, 1:20 serial fold dilutions of mouse sera were performed. Sera were incubated with SIVmac251.TCLA pseudovirus for 30 min in low-evaporation 96-well clear plates (Corning). TZM-bl cells were detached from flasks using 0.25% trypsin-EDTA (GIBCO) and seeded at a density of 0.5 × 106/ml per well. On the following day, 10% FBS complete DMEM (GIBCO) was added to each well. At day 3, media were aspirated, and cells were lysed using luciferase cell culture lysis buffer (Promega). Luciferase reaction was performed using 30 µl of cell lysis (Promega). The reaction was added to 96-well black optiplates (Perkin Elmer). Luminescence was measured using a Perkin Elmer Victor3 luminometer.
3
SARS-CoV-2 Spike Pseudovirus Neutralization Assay
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A SARS CoV-2 Spike pseudotyped lentivirus kit was obtained through BEI Resources, NIAID, NIH (SARS-Related Coronavirus 2, Wuhan-Hu-1 Spike-Pseudotyped Lentiviral Kit V2, NR-53816). We used Human Embryonic Kidney Cells (HEK-293T) expressing Human Angiotensin-Converting Enzyme 2, also known as HEK-293T-hACE2, which are susceptible to SARS CoV-2. This cell line was obtained through BEI Resources, NIAID, NIH NR-52511. Serial dilutions of sera were incubated with the SARS CoV-2 Spike pseudotyped lentivirus, following a protocol by Balazs and Bloom (20 ). Cells were lysed using luciferase cell culture lysis buffer (Promega). Luciferase reaction was performed using 30 μL of cell lysis (Promega). The reaction was added to 96-well black optiplates (Perkin Elmer). Luminescence was measured using Perkin Elmer Victor3 luminometer.
4
SARS-CoV-2 Spike Pseudovirus Neutralization Assay
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SARS-CoV-2 pseudovirus neutralization assays were performed as described previously.11 (link)
,49 (link) In brief, we utilized a SARS-CoV-2 spike pseudotyped lentivirus kit obtained through BEI Resources, NIAID, NIH (SARS-Related Coronavirus 2, Spike-Pseudotyped Lentiviral Kit V2, NR-53816). We used HEK-293T–hACE2 cells as targets (BEI Resources, NIAID, NIH, NR-52511). Serial plasma dilutions were incubated with the SARS-CoV-2 spike pseudotyped lentivirus (ancestral or Omicron BA.1). Cells were then lysed using luciferase cell culture lysis buffer (Promega). The reaction was added to 96-well black optiplates (PerkinElmer) and luminescence was quantified using a PerkinElmer Victor3 luminometer.
,49 (link) In brief, we utilized a SARS-CoV-2 spike pseudotyped lentivirus kit obtained through BEI Resources, NIAID, NIH (SARS-Related Coronavirus 2, Spike-Pseudotyped Lentiviral Kit V2, NR-53816). We used HEK-293T–hACE2 cells as targets (BEI Resources, NIAID, NIH, NR-52511). Serial plasma dilutions were incubated with the SARS-CoV-2 spike pseudotyped lentivirus (ancestral or Omicron BA.1). Cells were then lysed using luciferase cell culture lysis buffer (Promega). The reaction was added to 96-well black optiplates (PerkinElmer) and luminescence was quantified using a PerkinElmer Victor3 luminometer.
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