Bioluminometric assay of ATP in mouse brain: Determinant factors for enhanced test sensitivity.
Firefly luciferase bioluminescence (FLB) is a highly sensitive and specific method for the analysis of adenosine-5-triphosphate (ATP) in biological samples. Earlier attempts to modify the FLB test for enhanced sensitivity have been typically based on in vitro cell systems. This study reports an optimized FLB procedure for the analysis of ATP in small tissue samples. The results showed that the sensitivity of the FLB test can be enhanced several fold by using ultraturax homogenizer, perchloric acid extraction, neutralization of acid extract and its optimal dilution, before performing the assay reaction.
Khan
Haseeb Ahmad
HA
Department of Medical Research, Riyadh Al Kharj Hospital Program, Riyadh 11159, Kingdom of Saudi Arabia. khan_haseeb@yahoo.com
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000032:analysis* / Q000378:metabolism; D000818:Animals; D001671:Biochemistry / Q000379:methods*; D001921:Brain / Q000378:metabolism*; D001517:Coleoptera; D005260:Female; D049454:Firefly Luciferin / Q000378:metabolism; D008156:Luciferases / Q000378:metabolism; D008163:Luminescent Measurements; D051379:Mice; D008810:Mice, Inbred C57BL; D012680:Sensitivity and Specificity; D013997:Time Factors
D049454:Firefly Luciferin; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D008156:Luciferases