6′-Amino-D-luciferin 

6’-Amino-D-luciferin is a derivative of D-luciferin (HY-12591A) with an amino group replacing the 6-hydroxyl group. 6’-Amino-D-luciferin is a natural substrate of luciferase (Luc) and can be used for IVIS imaging in cell/animal experiments[1][2][3].

6′-Amino-D-luciferin (0.5 mg/mL; 2 min) red-shifts the bioluminescence spectrum of green-yellow emitting luciferase and blue-shifts the spectrum of red-emitting luciferase in a recombinant beetle luciferase assay, and the spectrum of the 5,5-dimethyl analog is almost identical to aminoluciferin^[2].
The total luminescence of 6′-Amino-D-luciferin (100 μM; 20 μL; 10-40 min) is approximately 74% of that of native luciferin^[2].
N-Z-DEVD-aLuc (1-100 μM; 45 min) increases caspase-3 activity in a dose-dependent manner; at 10 μM, it can be completely inhibited by Z-VAD-fmk (HY-16658B)^[3].
6′-Amino-D-luciferin can be used for IVIS imaging of cell/animal experiments to analyze the interaction of binding mutant active sites (λmax=588 nm).

1. Enzyme activity and kinetics assay
In vitro experiments^[1]:
Reaction system: 5-10 μL ATP/MgSO₄ mixture (40 mM/80 mM) + 10 μL enzyme solution (0.5-1 mg/mL) + 80-85 μL 0.5 mM 6′-Amino-D-luciferin (pH 8.0 Tris-HCl), luminescence intensity was detected at 22°C.
Cell experiment^[2]:
FLuc+ cells (MDA-MB-468): 6×10⁴ cells/well, 100 μM 6′-Amino-D-luciferin was added, and the luminescence kinetics were monitored using an IVIS200 imager (10 s integration, no filter).

2. Bioluminescence spectroscopy analysis
In vitro spectroscopy^[2]: 40 μL PBS + 20 μL ATP buffer (0.5 mM ATP, 0.5 mM CoA, 10 mM MgSO₄) + 1 μL substrate (1.5 mM) + 5 μL FLuc (130 μg/mL). FluoroMax-3 fluorescence instrument was used for acquisition (excitation light path blocked, 1 nm increment, 10 nm emission slit).
In vivo imaging^[2]: Nude mice were subcutaneously inoculated with FLuc+ tumor cells, and 2.5 mM 6′-Amino-D-luciferin (100 μL) was injected into the tail vein. IVIS200 imaging (30 s integration, no filter) was performed and tumor signals were normalized.

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

The 6′-Amino-D-luciferin derivative N-Z-DEVD-aLuc (50 mg/kg; intraperitoneal injection; single dose; 30 min after administration) could detect Ac-915-induced apoptotic signals in the U87-Luc glioma transplantation model of male SCID mice, and its bioluminescence intensity was higher than that of the control group^[3].

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

279.34

C₁₁H₉N₃O₂S₂

161055-47-6

Solid

Light yellow to brown

O=C([C@@H]1N=C(C2=NC3=CC=C(N)C=C3S2)SC1)O

Room temperature in continental US; may vary elsewhere.

* Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles.
Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month. When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.

• [1]. Viviani VR, et al. Bioluminescence of beetle luciferases with 6'-amino-D-luciferin analogues reveals excited keto-oxyluciferin as the emitter and phenolate/luciferin binding site interactions modulate bioluminescence colors. Biochemistry. 2014 Aug 19;53(32):5208-20  [Content Brief]

  [2]. Conley NR, et al. A selenium analogue of firefly D-luciferin with red-shifted bioluminescence emission. Angew Chem Int Ed Engl. 2012 Apr 2;51(14):3350-3.  [Content Brief]

  [3]. Kovács AK, et al. Synthesis of N-peptide-6-amino-D-luciferin Conjugates. Front Chem. 2018 Apr 19;6:120.  [Content Brief]