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Assay Method Information

Assay Name:  Anti-HIV Activity
Description:  Previously, a series of two-fold dilution on test samples were carried out in a 96-well plate (50 μL/well). Two plates were made for measuring anti-HIV activity and measuring cytotoxicity. For each agent, measurement in duplicate was carried out. An MT-4 cell suspension of 2.510^5/mL was dispensed at 100 μL/well onto the 96-well plate containing the test samples. An HIV virus solution was dispensed at 50 μL/well onto the 96-well plate containing the test samples and the cell. To the plate for measuring cytotoxicity, a culture solution was dispensed at 50 μL/well. It was mixed by a plate mixer, and then incubated in a CO2 incubator for 4 days. The 96-well plate incubated for 4 days was observed with the naked eye and under a microscope, and it was confirmed that there is no problem with virus proliferation and inhibition in the wells of the positive control and the negative control. Thirty microliters of a MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) solution was dispensed to each well.A reaction was allowed to occur in a CO2 incubator for an hour. From each well, 150 μL of the supernatant was removed such that cells are not sucked. One-hundred and fifty microliters of a cell lysis solution was added thereto, and then it was mixed well by a plate mixer until the whole cells are lysed. The mixed 96-well plates were measured by a microplate reader at two wavelengths, 560 nm/690 nm. Based on the following calculation formula, 50% HIV inhibition concentration (EC50) was calculated.
Affinity data for this assay
 

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Last update November 1, 2007
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