About Us
Since our founding in 1987, we have been developing and producing high-quality reagents and kits for luciferase-based assays, assisting customers worldwide in setting up their assays.
We sell directly to our customers and are supported by a wide network of distributors. Additionally, we are an OEM manufacturer for other ATP reagent lines.
BioThema is dedicated to the analytical applications of the firefly luciferase reaction. This versatile reaction can be used for various assays, including cell number quantification, cell proliferation, cytotoxicity, all enzymes and metabolites involved in ATP-forming or degrading reactions, reporter gene assays, immunoassays, rapid cell counting, and hygiene control.
Our facilities are conveniently located 25 minutes south of Stockholm, Sweden.
Our Technology
BioThema specializes in the analytical applications of the firefly luciferase reaction, offering a range of ATP reagents and standards for various assays. Our products enable precise measurement of ATP in cells, bacteria, and other biological samples, supporting research and diagnostics worldwide.
BioThema focuses on detecting cells, measuring enzyme activity, and more through the firefly luciferase reaction. In this process, luciferin is activated with ATP (adenosine triphosphate) and then oxidized to oxyluciferin and CO2. The oxyluciferin emits light as it returns to its ground state, which is measured using a luminometer. This method allows for accurate and versatile analytical applications in a variety of biological research and diagnostic contexts.
BioThema focuses on detecting cells, measuring enzyme activity, and more through the firefly luciferase reaction. In this process, luciferin is activated with ATP (adenosine triphosphate) and then oxidized to oxyluciferin and CO2. The oxyluciferin emits light as it returns to its ground state, which is measured using a luminometer. This method allows for accurate and versatile analytical applications in a variety of biological research and diagnostic contexts.
ATP + Luciferin + O2 → AMP + Pyrophosphate + Oxyluciferin + CO2 + light
In ATP assays, the light emission can be either constant or decay rapidly. Thus, there are stable light and flash-type ATP reagents based on the firefly luciferase reaction. With stable light reagents, a negligible proportion of ATP is consumed per minute, resulting in almost constant but low light emission. These are advantageous features when e.g coupling the firefly luciferase reaction with e.g metabolic ATP forming/depleting reactions, as small changes in the reaction rate of the coupled reaction can be detected at high ATP concentrations.
With flash-type reagents, the sensitivity to ATP is good and their ATP consumption is inevitable.These types of reagents feature rapidly decreasing light, but with an initially high peak light. The peak height and decay rate are both proportional to luciferase activity of the flash type reagent. These reagents are suitable for detection of low levels of ATP.
BioThema has developed three types of ATP reagents:
ATP Reagent SL
Reliable and Stable Light Emission
Description: ATP Reagent SL (Stable Light) is designed for continuous monitoring of ATP concentration in reaction mixtures where ATP is either formed or degraded. It has a decay rate of less than 1% per minute.. This reagent ensures that the light emission remains stable, allowing for precise and continuous measurements.
Applications: ATP Reagent SL is also suitable for measuring ATP in cultivated eukaryotic cells, with the assay’s linear range covering 1 to 50,000 cells. This broad range makes it versatile for various cell culture applications, providing consistent and reliable results.
ATP Reagent HS
High Sensitivity for Low-Level Bacterial Detection
Description: ATP Reagent HS (High Sensitivity) offers enhanced detection capabilities with a decay rate of approximately 10% per minute. This reagent has 20 times the activity compared to ATP Reagent SL, making it ideal for detecting low levels of bacterial ATP. Bacterial cells typically contain around 1 amol of ATP per cell, and ATP Reagent HS can detect as little as 10 amol ATP, equivalent to approximately 10 bacterial cells.
Applications: With a high-sensitivity luminometer, the linear detection range extends from 10 to 100,000 amol ATP. The reagent can be manually injected into samples, and the light signal measurement only takes a few seconds, ensuring quick and efficient bacterial detection.
ATP Reagent SS
Super-sensitive Detection for Minute ATP Levels
Description: ATP Reagent SS (Super Sensitive) is engineered for ultra-sensitive ATP detection, featuring a decay rate of around 235% per minute. The activity of this reagent 235 times higher than that of ATP Reagent SL. After one minute, only 10% of the initial light remains, thus requiring programmed dispensors to deliver the injection of reagent and standard into the sample in order to capture the signal effectively.
Applications: The detection limit for ATP Reagent SS is less than 1 amol ATP, making it capable of detecting ATP contaminations on pipette tips, microplates, and other surfaces. Due to its high sensitivity, this reagent should be used only when extremely low levels of ATP need to be measured.
The light signal produced in the firefly luciferase reaction and measured with a luminometer,is usually expressed in Relative Light Units (RLU). These units vary between instruments, reagents, sample matrices, and with temperature. Therefore, it is necessary to calibrate each assay by measuring the light signal before and after adding a known amount of ATP Standard.
BioThema provides a series of certified liquid-stable ATP Standards. These standards should be added in a small volume (10 µL) to avoid changing the reaction conditions. The amount of ATP added with the ATP standard should be at least 10 times the amount of ATP from the sample. Using ATP standards in this way compensates for all types of analytical interference, ensuring correct results.
This is not the case if an ATP standard curve is used, as there can be interference from the sample matrix and variations in sample temperature. The above ATP reagents and ATP standards are combined with various other reagents into several kits for special purposes.
These reagents include:
- ATP Eliminating Reagent for degrading extracellular ATP when determining intracellular ATP.
- Extractant B/S for releasing adenine nucleotides from bacterial and somatic cells.
- D-luciferin (free acid, potassium salt, and sodium salt) for in vivo imaging.
- Cell Lysing Reagent for lysing somatic cells while leaving bacterial cells unaffected.
- Luciferin Substrate and ATP Substrate for assays of firefly luciferase.
We also manufacture customized reagents and kits for other companies. For freeze-dried ATP reagents, the minimum order is 800 vials.