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measuring cell proliferation with SpectraMax M5, M2 and Gemini microplate readers using the CyQUANT Cell Proliferation Assay

Note: The CyQUANT kit includes a l bacteriophage DNA standard. The data shown in this application note were generated using this standard, prepared in serial dilutions. Depending upon the assay you are running, you may wish to use the kits DNA standard or another DNA that is more appropriate for your sample, prepared according to the instructions in the Molecular Probes Product Information sheet MP-7026.


Prepare the cells
Choose the appropriate set of culture and freezing instructions from Molecular Probes CyQUANT Product Information sheet for the type of cells (adherent or non-adherent) and growing conditions (in microplates or culture vessels). Freeze the cells at -70C. For this application note, adherent cells were trypsinized to detach them from the culture vessel, then pelleted and frozen. Samples containing known numbers of cells were used to demonstrate assay performance.


Prepare the cell samples, standard curve DNA samples and blanks
Step 1. Remove the previously prepared frozen cell pellets (or microplate) from -70C storage. Thaw the frozen cells at room temperature for a few minutes.

Step 2. Pre-treat the cells with RNase by adding 100 L 1X cell-lysis buffer containing 180 mM NaCl and 1 mM EDTA to each well and to four wells without cells (blanks). If using previously pelleted cells, resuspend each sample in 100 L and transfer to the wells of
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Normal human serum collected off the clot from healthy normal humans. Each donor unit is negative for anti-HIV 1&2, HIV-Ag, HCV, and HBsAg by FDA required tests...
Normal human AB serum, pooled...
sterile filtered, off the clot, all male donors, Total Protein: 4.5 to 8.5 g/dl...
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