0 0 PubChem PubChem assay example Cell Proliferation & Viability Assay A luciferase-based cell proliferation/viability assay end point kit was used as the readout for this assay. The kit measures the amount of ATP present in the microtiter plate well. If ATP is not present, the catalytic conversion luciferin into oxyluciferin is not possible, and no luminescence results. Since metabolically active cells produce ATP, the absence of ATP correlates with the presence of inviable cells. This campaign was run with doxorubicin, an antibiotic used as an anti-cancer drug, as the positive control. In this assay, doxorubicin had a 50% cytotoxicity concentration (CC50) of approximately 100 nM. All data reported was normalized on a per-plate basis to wells that contained cells in the presence of 4 micromolar doxorubicin (i.e. 100% inhibition). The assay was conducted in 1536-well format. A mathematical algorithm was used to determine active compounds. Two values were calculated: (1) the average percent inhibition of all compounds tested, and (2) three times their standard deviation. The sum of these two values was used as a cutoff value to determine which compounds were active. Any compound that exhibited greater %inhibition than the cutoff value was declared active. Possible artifacts in this assay include, but are not limited to, compounds that interfere with the luciferase reaction, absorb luminescence, or precipitate. Compounds that were active in this assay were subject to further follow up. Data from these assays will be uploaded to PubChem when available. A suspension of Jurkat cells (clone E6.1; ATCC Cat# TIB-152, Invitrogen Cat # K1045) in RPMI-1640 media containing 10% dialyzed fetal bovine serum, 0.1mM NEAA, 1mM Sodium Pyruvate, 25mM HEPES, 5mM L Glutamine, and 1x antibiotic was prepared prior to assay. The concentration of cells in the suspension was 1 million/milliliter. The assay began by dispensing 5 microliters of cell suspension to each test well of a 1536 well plate. Then 20nL of test compound or control doxorubicin was added. Plates were lidded and incubated at 37◦ C at 5% CO2 for 48 hrs prior to the addition of 5ul/well of Cell-Titer Glow (Promega Corp., Madison Wisconsin). After ten minutes of incubation at room temperature, plates were read on the Viewlux Imaging UHTS reader (PerkinElmer Lifesciences, Turku, Finland). Percent inhibition was calculated from the median of the positive control doxorubicin at 4 micromolar final screening concentration. ######### This is an example of the assay with SIDs defined in PUBCHEM_SID column and with no REGIDs in PUBCHEM_EXT_DATASOURCE_REGID All XRefs are for demonstrative purposes only and don't reflect the assay description ######### http://www.yoursite.com/database.html 16421977 example of using PubMed ID http://www.yoursite.com/assay.html 10116 example of using taxonomy ID for Rattus norvegicus 1 INHIBITION %Inhibition relative to cells in the presence of 4 micromolar doxorubicin 1 15