MTT-Test[1]                               19.09.2001

MTT Stock Solution                                                         4°C    

25mg     MTT                              Sigma M-5655
[3-(4,5-dimethyldiazol-2-yl)-2,5 diphenyl Tetrazolium Bromid]

ad 5ml PBS, pH 7.4, filter sterile

Lysis Solution                                                                   RT

99.4ml   DMSO

0.6ml    Acetic acid or HCl

10g      SDS

Method

Change Medium

Add 100µl MTT Stock Solution per 1ml medium

Incubate 2-4h in CO2 incubator

Aspirate supernatant carefully, do not wash with PBS

Add equal volume lysis solution to every sample

Lysate over 5min, mix well

Measurement

End point measurement

570nm    test wavelength

630nm    reference wavelength

 

 


[1]T. Lindl and J. Bauer, Zell- und Gewebekultur. Einführung in die Grundlagen sowie ausgewählte Methoden und Anwendungen. 1994: 250.

T. Mosmann, Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Meth, 1983. 65: 55-63.