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.

Similar to strandard luciferine substrate, 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 (i.e., at a final concentration of 13.6 µM).

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 Q-103 and Q-108, derived from Hikarazine Z103 and Hikarazine 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, the final blood concentration should be around 13.6 µM, which corresponds to injecting 80 µL of Z103 or Z108 at 1.35 mM per mouse (assuming an average blood volume of 8 mL).

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.