TZM-bl HIV infectivity was tested following an established protocol35 (link). Briefly, the collected cell culture supernatants were loaded into 96-well plates containing 50,000 TZM-bl cells per well in RPMI 1640 medium with 10% FBS, and incubated at 37 °C for 48 h. After washing, the incubated TZM-bl cells were measured by Beta-Glo assay kit (Promega, Madison, WI).
Beta glo assay kit
Manufactured by Promega
Sourced in United States
The Beta-Glo assay kit is a luminescent-based detection system designed to quantify the activity of beta-galactosidase in cell samples. The kit contains the necessary reagents to perform the assay, including the substrate and the detection solution. The luminescent signal produced is proportional to the amount of beta-galactosidase present in the sample.
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Lab products found in correlation
4 protocols using beta glo assay kit
1
TZM-bl Cell-based HIV Infectivity Assay
2
Cellular Senescence Luminescence Assay
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Senescence was measured with a Beta-Glo Assay kit (Promega), according to the manufacturer’s instructions. Luminescence intensity was determined as relative light units (RLUs) using a GloMax-Multi Detection System: Luminometer (Promega, Madison, WI, USA).
3
Senescence Measurement using Beta-Glo Assay
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Senescence was measured with a Beta‐Glo Assay kit (Promega), according to the manufacturer's instructions. Luminescence intensity was determined as RLUs using a GloMax‐Multi Detection System: Luminometer (Promega, USA).
4
Luminescence-based Parasite Growth Assay
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An assay to determine whether the compounds affect parasite growth was performed as described elsewhere [8 (link)]. The multiplicity of infection (MOI) was 1:5 (parasite/host cell ratio). In brief, the test compounds were reconstituted in culture medium at five concentrations (between 0 – 1.5 µg/mL), and incubated with newly purified parasites in monolayers of HFF cells grown in 96-well optical bottom plates (Nunc; Fisher Scientific, Pittsburgh, USA). The cells treated with medium only served as the negative drug control, while the medium-only well was used for background signal correction. To validate the assay, pyrimethamine was included as a positive drug control. After a 72-h incubation at 37oC in a 5% CO2 atmosphere, the parasite viability was determined by luminescence using a Beta-Glo assay kit (Promega, Madison, USA). Three independent biological experiments were performed, and each of which had three tests.
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