Introduction

The three primary analogues of coelenterazine—Coelenterazine, Coelenterazine h, and Coelenterazine 400a—can all be employed in similar experimental settings. However, depending on the application, some analogues may yield more optimal results than others.

Additional Information

Coelenterazine (Aladdin Catalog C131248)

Commonly referred to as Native Coelenterazine, this compound is widely utilized as a substrate for Renilla luciferases (Rluc) and Gaussia luciferase (Gluc). It exhibits an emission spectrum between 460–470 nm and serves multiple applications, including:

· Calcium ion detection in live cells

· Gene reporter assays

· BRET (bioluminescence resonance energy transfer) studies

· ELISA

· Detection of superoxide anions in cells

Coelenterazine h (Not currently available)

While Coelenterazine h is less effective when used with Renilla or Gaussia luciferases, it performs better in in vitro applications involving calcium-activated photoproteins. Notably, it offers a 10- to 20-fold increase in luminescent intensity compared to native Coelenterazine, making it especially valuable for detecting subtle changes in calcium ion concentrations. Its emission remains within the 460–470 nm range.

Coelenterazine 400a (Aladdin Catalog C332017 )

This analogue is the preferred substrate for Rluc in BRET experiments. Coelenterazine 400a emits between 390–400 nm, which reduces spectral overlap and minimizes interference with GFP acceptor emissions, enhancing the accuracy of energy transfer measurements.

References

Thomson, C. M., Herring, P. J., & Campbell, A. K. (1998). The widespread occurrence and tissue distribution of the imidazolopyrazine luciferins. Journal of Bioluminescence and Chemiluminescence, 12(2), 87–91.

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