BioThema AB applications

Methodology: All ATP producing and ATP degrading enzymatic reactions can be continuously monitored by measuring the light from firefly luciferase/luciferin being present in the reaction mixture. Examples include protein kinases⁶⁷, adenylate kinase¹⁴, pyruvate kinase⁶, pyruvate phosphate dikinase, glycerol kinase^{43, 44}, creatine kinase isoenzymes^{8, 10, 18, 22}, ATPases¹⁴, ATP synthetases⁴⁹, aminoacyl tRNA synthetase. Substrates to such enzymes can be measured kinetically or with endpoint assays. Examples include ADP, AMP, pyrophosphate. When continuously monitoring ATP the luciferase ATP depletion should be negligible compared to the enzymatic reactions being measured.

BioThema is constantly getting requests about developing customized assays and we have been doing just that for more than 40 years. Usually is a matter of setting up new assay protocols for our already available kits but sometimes there’s a need for new reagents also. BioThema is a long time OEM manufacturer of ATP reagent and would be glad to help you make an assay that fits your need.

Methodology: All biological residues contain ATP. When living cells die ATP is degraded to ADP and finally AMP. Measurement of ATP+AMP is a proof of biological residues being present. This can be done using the unique Kikkoman LuciPac Pen (surfaces) or LuciPac Pen Aqua (liquids) with a portable instrument PD-30. Hygiene control measuring ATP only may underestimate the contamination if ATP has been degraded in the biological residues. On the other hand ATP is an indicator that the residues are still alive. Neither ATP nor ATP+AMP are a reliable measure of bacteria as these contain much less ATP than cells from animals or plants.

Methodology: Antibodies or antigens may be labelled with luciferase or ATP producing enzymes.  Biotinylated recombinant luciferase can be detected at 10^-20 mol and biotinylated acetate kinase at 10-^21 mol.

In biochemistry, molecular biology and biology there is a need to measure cells, enzymes, metabolites and DNA/RNA. The luc gene coding for firefly luciferase is a reliable in vivo or in vitro reporter gene. ATP (adenosine triphosphate) participates in a large number of these reactions. Using the firefly luciferase reaction emitting a light proportional to the ATP concentration makes it possible to follow the reactions in a simple and highly sensitive way. Calibration of the assays with our certified liquid-stable ATP standard provides robust and reliable results. Please have a look at the application listing below to navigate to the product that matches your need.