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Mtt assay protocol for peptide/protein

mtt assay prtocol

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#1 ramadavid

ramadavid

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Posted 04 September 2013 - 07:57 PM

Dear all,
I am novice to the cell culture and mtt assay.
I want to set my protocol to do mtt assay of peptide and protein.
I have some confused suggestion on medium that it should contain FBS or medium should free from FBS for incubation of peptide drug with cell from expert friends.
one group is saying that medium containing peptide for incubation with cell should not contain the FBS ( Fetal Bovine serum ). Incubate cell in these medium with peptide drug for 4 hr. Then change the cell medium with medium containing FBS. Incubate requisite time, then add mtt to check cyototoxicity.
Another export group saying, use the complete medium (with FBS ) containing peptide. Incubate the cell in these medium for requisite time. Then add mtt to check cytotoxicity. 

The both group are agree for the one point that for non-peptide drug one should use the complete medium ( with FBS ) and drug, incubate for requisite time, add mtt to check cytotoxicity. 

Please help to solve the anomality with the peptide  drugs. 

Thank you 






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