Diphenylterazine (DTZ) (Synonyms: DTZ)

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

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 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

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.

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.