Diphenylterazine (DTZ)
(Synonyms: DTZ) 目录号 : GC33428
Diphenylterazine (DTZ) 是 CTZ 的类似物，是一种很好的荧光素酶底物。
Cas No.:344940-63-2
Sample solution is provided at 25 µL, 10mM.
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Quality Control & SDS
- View current batch:
- Purity: >98.00%
- COA (Certificate Of Analysis)
- SDS (Safety Data Sheet)
- Datasheet
| Cell experiment [1]: | |
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Cell lines |
HEK 293T |
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Preparation Method |
Cell viability was determined using RealTime-Glo MT Cell Viability Assay (Promega) after incubation of HEK 293T cells with individual luciferin substrates for 24 h at 37 °C. Cell morphology was further evaluated using microscopy. |
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Reaction Conditions |
24h 30 μM Diphenylterazine |
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Applications |
Compared to other substrates，As a novel luciferase substrate based on CTZ analogue, Diphenylterazine has bright red-shift bioluminescence. Moreover, Diphenylterazine alone yielded very little background and thus exhibited excellent signal-to-background ratios , Furthermore, Diphenylterazine elicited minimal cell toxicity at millimolar concentrations. |
| Animal experiment [1]: | |
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Animal models |
BALB/c mice |
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Preparation Method |
Bioluminescence imaging of intravenously injected HEK 293T cells：BALB/c mice are used and transfected with cells expressing biotinase gene by injecting cells into the tail vein of BALB/c mice. After the diminishing of the bioluminescence, 0.3 µmol Diphenylterazine is intraperitoneally injected. Mice are imaged with a 1-min exposure per frame over a course of 20 min. |
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Dosage form |
0.3 µmol Diphenylterazine |
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Applications |
Diphenylterazine was used as an emerging luciferase reporter substrate in mice. The imaging of deep tissue targets in mice was evaluated by injecting luciferase gene expressing HEK 293T cells through the tail vein. First, Diphenylterazine as a substrate has no background in control mice, and when injected into mice transfected with the luciferase gene, the fluorescence emitted by Diphenylterazine can be detected, and the intensity of Diphenylterazine has increased significantly compared with other substrates. |
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References: [1]. Yeh HW, Karmach O, Ji A, Carter D, Martins-Green MM, Ai HW. Red-shifted luciferase-luciferin pairs for enhanced bioluminescence imaging. Nat Methods. 2017 Oct;14(10):971-974. doi: 10.1038/nmeth.4400. Epub 2017 Sep 4. PMID: 28869756; PMCID: PMC5678970. |
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Diphenylterazine (DTZ), an analog of CTZ, is a good luciferase substrate. Diphenylterazine is a bioluminescence agent. Diphenylterazine alone yields very little background, leading to excellent signal-to-background ratios. Furthermore, Diphenylterazine elicits minimal cell toxicity at millimolar concentrations. Diphenylterazine could be synthesized from inexpensive commercial reagents in two steps with excellent yields[1].
In cell experiments, we can see that Diphenylterazine has a lower signal-to-noise ratio than other substrates, and Diphenylterazine causes minimal cytotoxicity at milimolar concentrations.
In contrast, other tested substrate induced cell death within the tested substrate concentration range.
In the mouse experiment, the bioluminescence of the surface site, at 0.1 mM substrate concentration, Diphenylterazine brightness was higher than that of the control group.
The same was true for imaging of deep tissue targets, and Diphenylterazine injection into untransfected BALB/c mice did not produce any background emission, and fluorescence was still detectable when injected intravenously into mice[2].
References:
[1].Tian X, Zhang Y, Li X, Xiong Y, Wu T, Ai HW. A luciferase prosubstrate and a red bioluminescent calcium indicator for imaging neuronal activity in mice. Nat Commun. 2022 Jul 8;13(1):3967. doi: 10.1038/s41467-022-31673-x. PMID: 35803917; PMCID: PMC9270435.
[2]. Yeh HW, et al. Red-shifted luciferase-luciferin pairs for enhanced bioluminescence imaging. Nat Methods. 2017 Oct;14(10):971-974.
Diphenylterazine (DTZ) 是 CTZ 的类似物，是一种很好的荧光素酶底物。 Diphenylterazine 是一种生物发光剂。单独使用二苯基特拉嗪产生的背景非常小，从而获得出色的信噪比。此外，Diphenylterazine 在毫摩尔浓度下引起最小的细胞毒性。 Diphenylterazine可以采用廉价的商业试剂分两步合成，收率高[1]。
在细胞实验中，我们可以看到 Diphenylterazine 的信噪比低于其他底物，并且 Diphenylterazine 在毫摩尔浓度下引起的细胞毒性极小。
相比之下，其他受试底物在受试底物浓度范围内诱导细胞死亡。
在小鼠实验中，表面位点的生物发光，在底物浓度为0.1 mM时，二苯基四嗪的亮度高于对照组。
深部组织靶点成像也是如此，未转染的BALB/c小鼠注射二苯特拉嗪未产生背景发射，小鼠静脉注射仍可检测到荧光[2] .
| Cas No. | 344940-63-2 | SDF | |
| 别名 | DTZ | ||
| Canonical SMILES | O=C1C(CC2=CC=CC=C2)=NC3=C(C4=CC=CC=C4)NC(C5=CC=CC=C5)=CN31 | ||
| 分子式 | C25H19N3O | 分子量 | 377.44 |
| 溶解度 | DMF : 11 mg/mL | 储存条件 | Store at -20°C, protect from light |
| General tips | 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；一旦配成溶液，请分装保存，避免反复冻融造成的产品失效。 储备液的保存方式和期限：-80°C 储存时，请在 6 个月内使用，-20°C 储存时，请在 1 个月内使用。 为了提高溶解度，请将管子加热至37℃，然后在超声波浴中震荡一段时间。 |
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| Shipping Condition | 评估样品解决方案：配备蓝冰进行发货。所有其他可用尺寸：配备RT，或根据请求配备蓝冰。 | ||
| 制备储备液 | |||
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1 mg | 5 mg | 10 mg |
| 1 mM | 2.6494 mL | 13.2471 mL | 26.4943 mL |
| 5 mM | 0.5299 mL | 2.6494 mL | 5.2989 mL |
| 10 mM | 0.2649 mL | 1.3247 mL | 2.6494 mL |
| 第一步：请输入基本实验信息（考虑到实验过程中的损耗，建议多配一只动物的药量） | ||||||||||
| 给药剂量 | mg/kg |  |
动物平均体重 | g | |
每只动物给药体积 | ul | |
动物数量 | 只 |
| 第二步：请输入动物体内配方组成（配方适用于不溶于水的药物；不同批次药物配方比例不同，请联系GLPBIO为您提供正确的澄清溶液配方） | ||||||||||
| % DMSO % % Tween 80 % saline | ||||||||||
| 计算重置 | ||||||||||
计算结果:
工作液浓度： mg/ml；
DMSO母液配制方法： mg 药物溶于 μL DMSO溶液（母液浓度 mg/mL，
体内配方配制方法：取 μL DMSO母液，加入 μL PEG300，混匀澄清后加入μL Tween 80，混匀澄清后加入 μL saline，混匀澄清。
1. 首先保证母液是澄清的；
2.
一定要按照顺序依次将溶剂加入，进行下一步操作之前必须保证上一步操作得到的是澄清的溶液，可采用涡旋、超声或水浴加热等物理方法助溶。
3. 以上所有助溶剂都可在 GlpBio 网站选购。
ATP-Independent Bioluminescent Reporter Variants To Improve in Vivo Imaging
ACS Chem Biol2019 May 17;14(5):959-965.PMID: 30969754DOI: 10.1021/acschembio.9b00150
Coelenterazine (CTZ)-utilizing marine luciferases and their derivatives have attracted significant attention because of their ATP-independency, fast enzymatic turnover, and high bioluminescence brightness. However, marine luciferases typically emit blue photons and their substrates, including CTZ and the recently developed diphenylterazine (DTZ), have poor water solubility, hindering their in vivo applications. Herein, we report a family of pyridyl CTZ and DTZ analogs that exhibit spectrally shifted emission and improved water solubility. Through directed evolution, we engineered a LumiLuc luciferase with broad substrate specificity. In the presence of corresponding pyridyl substrates (i.e., pyCTZ, 6pyDTZ, or 8pyDTZ), LumiLuc generates highly bright blue, teal, or yellow bioluminescence. We compared our LumiLuc-8pyDTZ pair with several benchmark reporters in a tumor xenograft mouse model. Our new pair, which does not need organic cosolvents for in vivo administration, surpasses other reporters by detecting early tumors. We further fused LumiLuc to a red fluorescent protein, resulting in a LumiScarlet reporter with further red-shifted emission and enhanced tissue penetration. LumiScarlet-8pyDTZ was comparable to Akaluc-AkaLumine, the brightest ATP-dependent luciferase-luciferin pair, for detecting cells in deep tissues of mice. In summary, we have engineered a new family of ATP-independent bioluminescent reporters, which will have broad applications because of their ATP-independency, excellent biocompatibility, and superior in vivo sensitivity.
Development of an ATP-independent bioluminescent probe for detection of extracellular hydrogen peroxide
Org Biomol Chem2022 Aug 10;20(31):6231-6238.PMID: 35548907DOI: 10.1039/d2ob00436d
This work reports a new ATP-independent bioluminescent probe (bor-DTZ) for detecting hydrogen peroxide that is compatible with the Nanoluciferase enzyme. The probe is designed with an arylboronate ester protecting group appended to a diphenylterazine core via a self-immolative phenolate linker. Reaction with hydrogen peroxide reveals diphenylterazine, which can then react with Nanoluciferase to produce a detectable bioluminescent signal. Bor-DTZ shows a dose-dependent response to hydrogen peroxide and selectivity over other biologically relevant reactive oxygen species and can be applied to detect either intra- or extracellular species. We further demonstrate the ability of this platform to monitor fluxes in extracellular hydrogen peroxide in a breast cancer cell line in response to the anticancer treatment, cisplatin.
A caged imidazopyrazinone for selective bioluminescence detection of labile extracellular copper(ii)
Chem Sci2022 Mar 23;13(15):4352-4363.PMID: 35509459DOI: 10.1039/d1sc07177g
Copper is an essential redox-active metal that plays integral roles in biology ranging from enzymatic catalysis to mitochondrial respiration. However, if not adequately regulated, this redox activity has the potential to cause oxidative stress through the production of reactive oxygen species. Indeed, the dysregulation of copper has been associated with a variety of disease states including diabetes, neurodegenerative disorders, and multiple cancers. While increasing tools are being developed for illuminating labile intracellular copper pools and the trafficking pathways in which they are involved, significantly less attention has been given to the analogous extracellular labile pool. To address this gap, we have developed a bioluminescence-based imaging probe, picolinic ester caged-diphenylterazine (pic-DTZ) for monitoring labile, extracellular copper using a coelenterazine-like imidazopyrazinone and the genetically-engineered, marine-based luciferase, nanoluciferase. Unlike the more commonly-used firefly luciferase, nanoluciferase does not require ATP, allowing its application to the extracellular milieu. pic-DTZ demonstrates high metal and oxidation state selectivity for Cu(ii) in aqueous buffer as well as selectivity for labile pools over coordinatively inaccessible protein-bound Cu(ii). We demonstrate the potential of pic-DTZ as a diagnostic tool in human serum and plasma for copper-associated diseases. Additionally, we apply pic-DTZ to lend insight into the extracellular copper dynamic in anticancer treatments.