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) causes a red shift in the bioluminescence spectrum of green-yellow light-emitting luciferases and a blue shift in the spectrum of red light-emitting luciferases in recombinant beetle luciferase assays, while the spectrum of the 5,5-dimethyl analog is almost identical to that of aminoluciferin^[1].
6′-Amino-D-luciferin (100 μM; 20 μL; 10-40 min) produces a total luminescence output approximately 74% that of native luciferin^[2].
N-Z-DEVD-aLuc (1-100 μM; 45 min) increases caspase-3 activity; at 10 μM, this effect can be completely inhibited by Z-VAD-fmk (HY-16658B)^[3].
6′-Amino-D-luciferin is suitable for IVIS imaging in cell and animal experiments and can be used to analyze interactions with mutant active sites (λmax=588 nm).

Guidelines (The following is our recommended protocol. This protocol is provided for reference only and should be modified according to your specific needs).
1. Enzyme Activity and Kinetic Assays
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); bioluminescence intensity is measured at 22°C.
Cell experiments^[2]:
FLuc+ cells (MDA-MB-468): seed 6×10⁴ cells/well, add 100 μM 6′-Amino-D-luciferin, and monitor luminescence kinetics using an IVIS200 imaging system (10-second integration, no filter).

2. Bioluminescence Spectral 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). Spectra are collected using a FluoroMax-3 fluorometer (excitation light path blocked, 1 nm increments, 10 nm emission slit width).
In vivo imaging^[2]:
FLuc+ tumor cells are subcutaneously implanted into nude mice, followed by tail vein injection of 2.5 mM 6′-Amino-D-luciferin (100 μL). Imaging is performed using an IVIS200 system (30-second integration, no filter), and tumor signals are 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]