I'm trying to do an assay which uses different concentrations of poly-l-lysine on a glass-bottomed, 24-well-plate to see what the appropriate poly-l-lysine concentration is for cells to adhere. I am using a 0.01% solution of the polypeptide from Sigma-Aldrich, and their instructions entail using 1.0 ml per 25 cm^2. Given the 10 mm diameter of these wells, that works out to ~31.4 ul of the solution. Really? Isn't that awfully less? Has anyone else used this reagent and/or coated plates like so? If so, does anyone have any ideas/tips/tricks/things-to-avoid regarding this at all?
Thanks, y'all!
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This is probably not relevant anymore (I couldn't see the date this question was posted until I clicked on "Answer") and I guess the question was resolved eventually.
Anyways, you seem to have mistaken the diameter of the wells, which is ~15 - 16 mm for 24 well plates (15.6 mm as described at the link below). So that brings the volume to 76 µl, which should be enough to just cover the bottom.
Cheers!
http://www.sigmaaldrich.com/catalog/product/sigma/cls3527?lang=en®ion=US
Thanks, Nitesh! But, yeah, I crunched the numbers and figured it out eventually! :)
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