Bioluminescence FAQ

The choice between Hikarazine™ Z108 and Hikarazine™ Z103 depends on your specific assay.

• For applications where maximizing light output is crucial, such as bioluminescence microscopy and small animal imaging, Hikarazine™ Z108 is generally the best choice.
• For applications requiring longer monitoring times, Hikarazine™ Z103 is recommended. Although it is slightly less bright, some users also report an improved signal-to-noise ratio under specific conditions when using Hikarazine™ Z103.

The choice of buffer is crucial for optimal bioluminescence. PBS is recommended, but other buffers may be used depending on the assay. For instance, adding 0.01% Tween can help with complex matrices, though it may slightly increase background noise. Additionally, varying cation compositions can influence both brightness and bioluminescence half-life. If you need further guidance after testing in your specific assay, please contact our experts.

The light emission and brightness of Hikarazine™ are influenced by several factors:

• The amount of oxygen available at the luciferase site,
• The buffer composition, which can promote non-radiative relaxation of the substrate’s excited state,
• Temperature, which can affect luciferase activity and the stability of the enzyme-substrate complex, thus altering light intensity,
• Ion concentration, particularly divalent cations, which can influence the efficiency of the bioluminescence reaction,
• The pH of the medium, as inappropriate pH can alter the luciferase enzyme’s activity,
• The presence of cofactors or inhibitors, as some compounds can enhance the reaction’s efficiency, while others may reduce it.

These factors must be optimized to ensure the best performance of the bioluminescent system using Hikarazine™.

To ensure stability for up to two years, store the vials at room temperature, away from heat, light, and humidity.

A slight darkening of the surface of the solid may occur over time. However, this does not affect the bioluminescence signal, provided the pro-substrate is used under standard conditions recommended in the IFU.

The stock solution is acidic, which enhances its stability. When stored at 4°C, it remains stable for 6 months. For extended stability, it can be stored at -20°C, with ongoing tests to determine the exact duration.

The peak bioluminescence emission of activated luciferins Hikarazine™ Z103 and Z108 has been measured at 450 ± 3 nm.

Hikarazine™ substrates contain fluor atoms, enhancing their solubility and bioavailability for small animal imaging.
For optimal results, we recommend injecting 100 to 150 µL of Z103 or Z108 at a concentration of 1 mM (target dose of 3 mg/kg for mice weighing approximately 20 g).

No, unlike firefly-based bioluminescent systems, marine-based bioluminescent systems, including Hikarazine™, do not require ATP.

Yes, Hikarazine™substrates are patented under EP3615516, US11938199, and JP7366752.

Yes, Hikarazine™ substrates are compatible with most bioluminescence detection systems, including luminometers, plate readers and in vivo imaging systems, including Berthold Technologies instruments, as well as other major detection platforms.

No, thanks to the solubility of the substrates, Hikarazine™ can be added directly to the cell culture medium, allowing bioluminescence measurements to be performed in live cells. 

For the activated form of Hikarazine™ substrates, we recommend starting bioluminescence acquisitions 5 minutes after injection.