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

te lysis of the cells.


Prepare the assay reagents
Step 1. Dilute the concentrated cell-lysis buffer stock solution 1:20 with distilled water (prepare 200 L per well) to make 1X celllysis buffer.

Step 2. Prepare 1X working solution of CyQUANT GR dye/cell-lysis buffer by diluting the concentrated CyQUANT GR stock solution 1:400 with 1X cell-lysis buffer. This solution should be prepared in a plastic container rather than glass (see CyQUANT product insert).


Prepare cell-based standard curve and unknowns
Step 1. Thaw the previously prepared frozen cell pellet at room temperature for a few minutes. Add 1.0 mL of CyQUANT GR dye/cell-lysis buffer and resuspend the cells by vortexing briefly. Assuming the pellet contained 106 cells, the resulting cell lysate contains the equivalent of 106 cells/mL.

Step 2. Generate a dilution series in the wells of a microplate, starting at 50,000 cells/well and making serial 1:2 dilutions down to 24 cells/well. se CyQUANT GR dye/cell-lysis buffer to make dilutions such that the desired cell number is in a volume of 200 l. Prepare four replicates of each standard, and include a set of replicates without cells as a control (you can omit one of the standards, e.g., 781 cells/well, to make room on the microplate).

Step 3. Incubate the microplate at room temperature for 2-5 minutes, protected from light.


Set up the instrument and software
Step 1. Turn on the SpectraMax M5, M2, or Gemini microplate reader. Launch the SoftMax Pro software and use the Assays dro
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