Cytotoxicity assay with RAW 264.6 cells - (Sep/30/2008 )

Hi everybody!
Which is the best way to measure cytotoxicity with RAW cells? Realese of LDH enzyme?
Dr. UDPG

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LDH and MTT both work fine..............i have used both the methods and found both of them to give similar results. However, MTT had a plus point that you can handle several hundreds of samples (using 96- or 384- well plates) while for LDH you need to analyze each sample individually by an enzyme kinetic assay. The chromogenic assay of LDH (which is not a kinetic assay) can also be performed but variablity is high.

The selection of assay also depends on the type of material to test. The membrane damaging materials will give you high LDH values without much change in MTT assay. On the other hand, several compounds, while actually being toxic, may increase mitochondrial mass and may show no change in cell viability by MTT.

So, it is always better to use two assays together to determine cytotoxicity of compounds.

You may use assays that estimate DNA or proteins. Also, you can can quantify fluorescence of fluorescein diacetate to estimate cytotoxicity.

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