Home>>Natural Products>>Diphenylterazine (DTZ)

Diphenylterazine (DTZ)

Catalog No.: GC33428

Diphenylterazine (DTZ) (DTZ) is a bioluminescence agent.

Diphenylterazine (DTZ) Chemical Structure

Size Price Stock Qty
In stock
In stock
In stock
In stock

Customer Reviews

Based on customer reviews.

Tel: (626) 353-8530 Email: sales@glpbio.com

Sample solution is provided at 25 µL, 10mM.

Product Documents

Quality Control & SDS

View current batch:


Cell experiment [1]:

Cell lines

HEK 293T

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.

Reaction Conditions

24h 30 μM Diphenylterazine


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 [2]:

Animal models

BALB/c mice

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.

Dosage form

0.3 µmol Diphenylterazine


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.


[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.


Diphenylterazine, 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].

[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.

Chemical Properties

Cas No. 344940-63-2 SDF
Canonical SMILES O=C1C(CC2=CC=CC=C2)=NC3=C(C4=CC=CC=C4)NC(C5=CC=CC=C5)=CN31
Formula C25H19N3O M.Wt 377.44
Solubility DMSO : ≥ 19 mg/mL (50.34 mM);Water : < 0.1 mg/mL (insoluble) Storage Store at -20°C, protect from light
General tips For obtaining a higher solubility , please warm the tube at 37 ℃ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20℃ for several months.
Shipping Condition Evaluation sample solution : ship with blue ice
All other available size: ship with RT , or blue ice upon request

In vivo Formulation Calculator (Clear solution)

Step 1: Enter information below (Recommended: An additional animal making an allowance for loss during the experiment)

mg/kg g μL

Step 2: Enter the in vivo formulation (This is only the calculator, not formulation. Please contact us first if there is no in vivo formulation at the solubility Section.)

% DMSO % % Tween 80 % saline

Calculation results:

Working concentration: mg/ml;

Method for preparing DMSO master liquid: mg drug pre-dissolved in μL DMSO ( Master liquid concentration mg/mL, Please contact us first if the concentration exceeds the DMSO solubility of the batch of drug. )

Method for preparing in vivo formulation: Take μL DMSO master liquid, next addμL PEG300, mix and clarify, next addμL Tween 80, mix and clarify, next add μL saline, mix and clarify.

Method for preparing in vivo formulation: Take μL DMSO master liquid, next add μL Corn oil, mix and clarify.

Note: 1. Please make sure the liquid is clear before adding the next solvent.
2. Be sure to add the solvent(s) in order. You must ensure that the solution obtained, in the previous addition, is a clear solution before proceeding to add the next solvent. Physical methods such as vortex, ultrasound or hot water bath can be used to aid dissolving.
3. All of the above co-solvents are available for purchase on the GlpBio website.

  • Molarity Calculator

  • Dilution Calculator

**When preparing stock solutions always use the batch-specific molecular weight of the product found on the vial label and MSDS / CoA (available online).


Research Update

ATP-Independent Bioluminescent Reporter Variants To Improve in Vivo Imaging

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.

A caged imidazopyrazinone for selective bioluminescence detection of labile extracellular copper(ii)

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.

Development of an ATP-independent bioluminescent probe for detection of extracellular hydrogen peroxide

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.


Review for Diphenylterazine (DTZ)

Average Rating: 5 ★★★★★ (Based on Reviews and 4 reference(s) in Google Scholar.)

5 Star
4 Star
3 Star
2 Star
1 Star
Review for Diphenylterazine (DTZ)

GLPBIO products are for RESEARCH USE ONLY. Please make sure your review or question is research based.

Required fields are marked with *

You may receive emails regarding this submission. Any emails will include the ability to opt-out of future communications.