please send aprotocol for MMT assay

StewJW via methods%40net.bio.net (by stewjw from gmail.com)
Wed Mar 21 12:15:31 EST 2007


Here's an extract from one of my papers. The assay was done in a
mirotitre plate format:

In vitro XTT cytotoxicity Assay. The determination of cell viability
was accomplished by the addition of an XTT solution to J774.A1
cultures [54, 55] treated with graded concentrations of the test
compounds. Cell culture and plating procedures were performed as
described previously for nitric oxide measurement. J774.A1 cells in 96
well plates were treated with serially diluted concentrations of PMB,
or test compounds to a final volume of 200 μl/well and left to
incubate at 37°C for 16h. Wells containing J774.A1 medium only served
as the negative control. Cytotoxicity was measured the following day
by the addition of 80 μl/well XTT/Phenazine methosulfate (PMS)
solution (XTT solution, 2 mM in PBS, pH 7.4, pH adjusted to 6.0-6.5;
PMS solution, 0.92 mg/ml in PBS, pH 7.4; solutions mixed at a ratio of
8 ml XTT solution to 200 μl PMS solution) followed by an incubation
time of 1.5h at 37°C. Absorbance was read at 490 nm with scatter
correction at 690 nm..

The full paper which can be downloaded free is:
http://www.pubmedcentral.nih.gov/articlerender.fcgi?tool=pubmed&pubmedid=15578935

XTT is better than MTT.



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