Coelenterazine is a luminescent enzyme substrate for apoaequorin and Renilla luciferase. Renilla luciferase and substrate coelenterazine has been used as the bioluminescence donor in bioluminescence resonance energy transfer (BRET) to detect protein-protein interactions. Coelenterazine is a superoxide anion-sensitive chemiluminescent probe and it can also be used in chemiluminescent detection of peroxynitrite^[1][2][3].

HCT-8 control cells, transiently expressing Renilla luciferase (RLuc), showed low bioluminescence due to P-glycoprotein–mediated efflux transport of coelenterazine. By comparison, transiently expressing RLuc HCT-8 cells, wherein P-glycoprotein was down-regulated with shRNAi, showed high bioluminescence^[3].

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

The in vivo growth potential of HCC1806-RR was monitored by injecting animals with coelenterazine (2 mg/kg) i.v. and exposing them to a charged-coupled device (CCD) camera 5 minutes later. Rluc activity was detected as light emitted from the tumor cells and acquired as a pseudo-color image superimposed over a black and white photograph of the animal. All mice demonstrated very high Rluc activity at the primary site with the majority of mice simultaneously showing metastases to inguinal ILNs^[4].

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

423.46

C₂₆H₂₁N₃O₃

55779-48-1

腔肠素

Room temperature in continental US; may vary elsewhere.

-20deg;C, protect from light, stored under nitrogen

*该产品在溶液状态不稳定，建议您现用现配，即刻使用。

Ethanol : 2 mg/mL (4.72 mM; Need ultrasonic; DMSO can inactivate Coelenterazine’s activity)

请根据产品在不同溶剂中的溶解度，选择合适的溶剂配制储备液；该产品在溶液状态不稳定，建议您现用现配，即刻使用。

请根据您的实验动物和给药方式选择适当的溶解方案。以下溶解方案都请先按照 In Vitro 方式配制澄清的储备液，再依次添加助溶剂：

——为保证实验结果的可靠性，澄清的储备液可以根据储存条件，适当保存；体内实验的工作液，建议您现用现配，当天使用； 以下溶剂前显示的百
分比是指该溶剂在您配制终溶液中的体积占比；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的方式助溶

• 1.

  请依序添加每种溶剂：;10% EtOH ;; 40% PEG300 ;; 5% Tween-80 ;; 45% saline

  Solubility: ≥ 0.2 mg/mL (0.47 mM); Clear solution

  此方案可获得 ≥ 0.2 mg/mL (0.47 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 2.0 mg/mL 的澄清 EtOH 储备液加到 400 μL PEG300 中，混合均匀；向上述体系中加入50 μL Tween-80，混合均匀；然后继续加入 450 μL生理盐水定容至 1 mL。

  将 0.9 g 氯化钠，完全溶解于 100 mL ddH₂O 中，得到澄清透明的生理盐水溶液

• 2.

  请依序添加每种溶剂：;10% EtOH ;; 90% (20% SBE-β-CD in saline)

  Solubility: ≥ 0.2 mg/mL (0.47 mM); Clear solution

  此方案可获得 ≥ 0.2 mg/mL (0.47 mM，饱和度未知) 的澄清溶液。

  以 1 mL 工作液为例，取 100 μL 2.0 mg/mL 的澄清 EtOH 储备液加到 900 μL 20% 的 SBE-β-CD 生理盐水水溶液中，混合均匀。

  将 2 g 磺丁基醚 β-环糊精加入 5 mL 生理盐水中，再用生理盐水定容至 10 mL，完全溶解，澄清透明
• 3.

  请依序添加每种溶剂：;10% EtOH ;; 90% Saline

  Solubility: ≥ 0.2 mg/mL (0.47 mM); Clear solution

• [1]. Markova SV, et al, Coelenterazine-dependent luciferases. Biochemistry (Mosc). 2015 Jun;80(6):714-32.

  [2]. Lucas M, et al. Coelenterazine is a superoxide anion-sensitive chemiluminescent probe: its usefulness in the assay of respiratory burst in neutrophils. Anal Biochem. 1992 Nov 1;206(2):273-7.

  [3]. Pichler A, et al. In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein. Clin Cancer Res. 2005 Jun 15;11(12):4487-94.

  [4]. Volk-Draper LD, et al. Novel model for basaloid triple-negative breast cancer: behavior in vivo and response to therapy. Neoplasia. 2012 Oct;14(10):926-42.

Cells are plated at a density of 5×10⁴ cells per well into 24-well plates and grown to 80-100% confluency. Just before imaging, media are changed to a colorless solution containing (in mM): 2.7 KCl, 139 NaCl, 8.1 Na₂HPO₄, 0.7 H₂O, 1.5 KH₂PO₄, 1.8 CaCl₂, 1 MgCl₂ and 5.5 d-glucose. Cells are preincubated for 15 min in the absence or presence of Pgp modulator, after which Coelenterazine (final concentration of 470 nM) is added directly to the cells^[1].

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

Mice are anesthetized with metofane before tail vein injection of Coelenterazine (4 μg/g) formulated from an ethanol stock diluted in sodium phosphate buffer (50 mM). Bioluminescence imaging is performed on the in vivo imaging system at 2, 6, 8, and 11 min after injection. After imaging, animals are killed by cervical dislocation; tumors are then harvested and weighed^[1].

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

• [1]. Markova SV, et al, Coelenterazine-dependent luciferases. Biochemistry (Mosc). 2015 Jun;80(6):714-32.

  [2]. Lucas M, et al. Coelenterazine is a superoxide anion-sensitive chemiluminescent probe: its usefulness in the assay of respiratory burst in neutrophils. Anal Biochem. 1992 Nov 1;206(2):273-7.

  [3]. Pichler A, et al. In vivo RNA interference-mediated ablation of MDR1 P-glycoprotein. Clin Cancer Res. 2005 Jun 15;11(12):4487-94.

  [4]. Volk-Draper LD, et al. Novel model for basaloid triple-negative breast cancer: behavior in vivo and response to therapy. Neoplasia. 2012 Oct;14(10):926-42.