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] .