Serum albumin leads to false-positive results in the XTT and the MTT assay

BioTechniques

Volumn 43, Issue 2, 2007, Pages 178-186

  Funk, Dorothee ^a   Schrenk, Hans Hermann ^a   Frei, Eva ^a  

^a GERMAN CANCER RESEARCH CENTER DKFZ   (Germany)

Author keywords

[No Author keywords available]

Indexed keywords

POTENTIAL CYTOTOXIC EFFECTS; PROTEOLYTIC DIGESTION; SERUM ALBUMIN; TETRAZOLIUM SALTS;

BIOASSAY; CELLS; CYTOTOXICITY; REDUCTION; SALTS;

ENZYME KINETICS;

3 (4,5 DIMETHYL 2 THIAZOLYL) 2,5 DIPHENYLTETRAZOLIUM BROMIDE; BENZENE DERIVATIVE; BOVINE SERUM ALBUMIN; CYSTEINE; FORMAZAN; GLUTATHIONE; HUMAN SERUM ALBUMIN; N ETHYLMALEIMIDE; NITRO DERIVATIVE; SERUM ALBUMIN; SODIUM 2,3 BIS(2 METHOXY 4 NITRO 5 SULFOPHENYL) 2H TETRAZOLIUM 5 CARBOXANILIDE; SODIUM DERIVATIVE; SULFONE DERIVATIVE; TETRAZOLE DERIVATIVE; UNCLASSIFIED DRUG;

CELL COUNT; CELL VIABILITY; CONCENTRATION RESPONSE; CYTOTOXICITY; ENZYME ACTIVITY; PROTEIN DEGRADATION; REACTION ANALYSIS; REDUCTION; REVIEW;

ARTIFACTS; BIOLOGICAL ASSAY; CELL COUNT; CELL-FREE SYSTEM; FALSE POSITIVE REACTIONS; FORMAZANS; HUMANS; REPRODUCIBILITY OF RESULTS; SENSITIVITY AND SPECIFICITY; SERUM ALBUMIN; TETRAZOLIUM SALTS;

BOVINAE;

EID: 34547879316     PISSN: 07366205     EISSN: None     Source Type: Journal    
DOI: 10.2144/000112528     Document Type: Review

References (16)

1. Mosmann, T. 1983. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J. Immunol. Methods 65:55-63.
2. Marshall, N.J., C.J. Goodwin, and S.J. Holt. 1995. A critical assessment of the use of microculture tetrazolium assays to measure cell growth and function. Growth Regul. 5:69-84.
3. Goodwin, C.J., S.J. Holt, S. Downes, and N.J. Marshall. 1995. Microculture tetrazolium assays: a comparison between two new tetrazolium salts, XTT and MTS. J. Immunol. Methods 179:95-103.
4. Hamasaki, K., K. Kogure, and K. Ohwada. 1996. A biological method for the quantitative measurement of tetrodotoxin (TTX): tissue culture bioassay in combination with a water-soluble tetrazolium salt. Toxicon 34:490-495.
5. Roehm, N.W., G.H. Rodgers, S.M. Hatfield, and A.L. Glasebrook. 1991. An improved colorimetric assay for cell proliferation and viability utilizing the tetrazolium salt XTT. J. Immunol. Methods 142:257-265.
6. Slater, T.F., B. Sawyer, and U. Straeuli. 1963. Studies on succinate-tetrazolium reductase system. III. Points of coupling of four different tetrazolium salts. Biochim. Biophys. Acta 77:383-393.
7. Pagliacci, M.C., F. Spinozzi, G. Migliorati, G. Fumi, M. Smacchia, F. Grignani, C. Riccardi, and I. Nicoletti. 1993. Genistein inhibits tumour cell growth in vitro but enhances mitochondrial reduction of tetrazolium salts: a further pitfall in the use of the MTT assay for evaluating cell growth and survival. Eur. J. Cancer 29A:1573-1577.
8. Jabbar, S.A., P.R. Twentyman, and J.V. Watson. 1989. The MTT assay underestimates the growth inhibitory effects of interferons. Br. J. Cancer 60:523-528.
9. Natarajan, M., S. Mohan, B.R. Martinez, M.L. Meltz, and T.S. Herman. 2000. Antioxidant compounds interfere with the 3-[4,5- dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide cytotoxicity assay. Cancer Detect. Prev. 24:405-414.
10. Chakrabarti, R., S. Kundu, S. Kumar, and R. Chakrabarti. 2000. Vitamin A as an enzyme that catalyzes the reduction of MTT to formazan by vitamin C. J. Cell. Biochem. 80:133-138.
11. Bernhard, D., W. Schwaiger, R. Crazzolara, I. Tinhofer, R. Kofler, and A. Csordas. 2003. Enhanced MTT-reducing activity under growth inhibition by resveratrol in CEM-C7H2 lymphocytic leukemia cells. Cancer Lett. 195:193-199.
12. Huang, K.T., Y.H. Chen, and A.M. Walker. 2004. Inaccuracies in MTS assays: major distorting effects of medium, serum albumin, and fatty acids. BioTechniques 37:406-412.
13. Weigand, M., G. Hartung, J. Roboz, S. Sieger, M. Wolf, H. Sinn, H.H. Schrenk, M. Wiessler, and E. Frei. 2001. Mode of action of methotrexate-albumin in a human T-cell leukemia line and activity against an MTX-resistant clone. Anticancer Drug Des. 16:227-237.
14. Van Noorden, C.J., A. Kooij, I.M. Vogels, and W.M. Frederiks. 1985. On the nature of the 'nothing dehydrogenase' reaction. Histochem. J. 17:1111-1118.
15. Peters, T. 1996. All About Albumin: Biochemistry, Genetics and Medical Applications. Academic Press, San Diego.
16. Zhang, Z.D. and G. Cox. 1996. MTT assay overestimates human airway smooth muscle cell number in culture. Biochem. Mol. Biol. Int. 38:431-436.