Coelenterazine h

Coelenterazine h is a dehydroxy derivative of natural coelenterazine and serves as a luminescent substrate for RLuc8. Coelenterazine h is Ca²⁺-sensitive, making it an effective tool for measuring minute changes in Ca²⁺ concentration. Coelenterazine h is widely used in cell biology, molecular biology, drug discovery, and in vivo imaging.

superoxide anion (Rat Hepatocyte cells):69 μM

Methods: In BRET sensor (BRAC) studies, Coelenterazine-h (20 μM in vitro, 10 μM in cells) serves as the substrate for RLuc8, initiating the BRET signaling chain for real-time monitoring of Ca²⁺ binding kinetics. Changes in Venus (530 nm) emission were recorded at a high frequency of 1 kHz via stop-and-go spectroscopy.
Results: The Ca²⁺ dissociation time constant τ_off = 0.21 s was successfully resolved, approximately 14 times faster than conventional FRET indicators. [1]
Methods: C6 (rat glioma) cells were transfected with pCMV-Rluc plasmid. Cell lysates were obtained 48 hours post-transfection. Cell lysates were mixed with Coelenterazine h (0.1, 1, 10, 50, 100, 200 μg/mL) and measured for 10 seconds. A luminometer recorded RLU/mg protein.
Results: Luminescence increased with Coelenterazine h concentration, peaking at 50 μg/mL, then decreased at higher doses. [2]
Methods: Human breast cancer cells (MCF-7) were treated with Rluc8.6-KR-LP (10 μM) for 24 hours, followed by Coelenterazine h (150 μM) for an additional 24 hours. Analysis included fluorescence microscopy (DAPI staining for live cells, SYTOX Green for dead cells, KR red fluorescence tracing) and flow cytometry quantification.
Results: The Rluc8.6-KR-LP + Coelenterazine H group exhibited substantial cell death, with KR red fluorescence localized to the cell membrane. [3]

Methods: C6-Rluc cells (1×10⁶) were injected into the mouse tail vein (cells naturally homed to liver/lung). Coelenterazine h (2.8 mg/kg) was administered via tail vein injection 90 minutes after cell injection. Imaging was performed using a cooled CCD camera for 7-10 minutes. Following euthanasia, tissue homogenates were collected and verified using a luminometer.
Results: Signals were detectable in the chest and upper abdomen (liver/lung regions), peaking at 5 minutes and decaying between 7–10 minutes. Tissue homogenate analysis confirmed signals primarily originated from the lungs. [2]
Methods: NOD SCID-γ immunodeficient mice subcutaneously implanted with MDA-MB-231 human breast cancer cells. Three intra-tumoral injections of Rluc8.6-KR-LP (1 nmol) were administered via intratumoral injection. Coelenterazine h (5 μg) was injected subcutaneously on the second day after protein injection. Tumor volume was measured 2-3 times weekly.
Results: Rluc8.6-KR-LP + Coelenterazine h significantly inhibited tumor growth, with an average volume of 127 mm³ on day 34.[3]

DMSO: 237.5 mg/mL (582.88 mM), Sonication is recommended.