Coelenterazine h  (Synonyms: 2-Deoxycoelenterazine; CLZN-h)

Coelenterazine h (2-Deoxycoelenterazine), a coelenterazine derivative, is a luminescent substrate for RLuc8. Coelenterazine h is more sensitive to Ca²⁺, thus providing a valuable tool for measuring small changes in Ca²⁺ concentrations (Ex/Em = 437/466 nm)^[1][2][3][4].

Coelenterazine h (at concentrations of 1-10 μM) can be used as the luminescent substrate for RLuc8 [4].
In the Ca²⁺ binding kinetics measurement of BRAC, 5 nM BRAC protein was rapidly mixed with 20 μM Coelenterazine h in different concentrations of Ca²⁺ buffer solution, and the Venus (530 nm) emission intensity of BRAC was monitored at 1 kHz [4].
Guide (The following is our recommended solution. This solution is merely a guideline and should be modified according to your specific needs.)
1. Solution Preparation^[4]
1.1 Preparation of Stock Solution
Solvent: Methanol or Ethanol
Concentration: 10 mM (optimized according to the experiment).
1.2 Solution Preparation
Dilute to 10 μM using PBS or cell culture medium (based on experimental optimization).
Note: The raw solution and working solution should be prepared and used immediately. Store them in a dark place.

2. Cell Staining
2.1 Transfect Hela cells with BRAC or G5A.
2.2 Add 10 μM Coelenterazine h to the cell plate.
2.3 Add the dye and the template to the cell culture box and incubate for 1 to 4 hours (depending on the experimental optimization).
2.4 Fluorescence microscopy detection (Ex/Em = 437/466 nm).

MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.

407.46

C₂₆H₂₁N₃O₂

50909-86-9

Solid

Yellow to orange

OC(C=C1)=CC=C1C2=CN(C(C(CC3=CC=CC=C3)=N4)=O)C4=C(N2)CC5=CC=CC=C5

Room temperature in continental US; may vary elsewhere.

-20°C, sealed storage, away from moisture and light

*The compound is unstable in solutions, freshly prepared is recommended.

• [1]. Jiang T, et al. New bioluminescent coelenterazine derivatives with various C-6 substitutions. Org Biomol Chem. 2017 Aug 23;15(33):7008-7018.  [Content Brief]

  [2]. M R Knight, et al. Imaging calcium dynamics in living plants using semi-synthetic recombinant aequorins. J Cell Biol. 1993 Apr;121(1):83-90.  [Content Brief]

  [3]. Kazushi Suzuki, et al. Five colour variants of bright luminescent protein for real-time multicolour bioimaging. Nat Commun. 2016 Dec 14:7:13718.  [Content Brief]

  [4]. Kenta Saito, et al. Auto-luminescent genetically-encoded ratiometric indicator for real-time Ca2+ imaging at the single cell level. PLoS One. 2010 Apr 1;5(4):e9935.  [Content Brief]