AkaLumine

AkaLumine is a D-luciferin (HY-12591A) analogue with a K_m of 2.06 μM for recombinant firefly luciferase (Fluc) protein. AkaLumine emits near-infrared (NIR) light (λmax=677 nm) in reactions with native Fluc. AkaLumine has high tissue-penetration and increases detection sensitivity from deep-tissue targets^[1]. IC50 & Target:Km: 2.06 μM^[1]. In Vitro:1. Solution preparation^[2]
1.1 Preparation of stock solution
Solvent: Anhydrous DMSO or ultrapure water.
Concentration: 10 mM is recommended (e.g., weigh 3.17 mg AkaLumine hydrochloride and dissolve in 1 mL solvent).
Storage: Store at -20°C in dark after aliquoting and avoid repeated freezing and thawing.
1.2 Preparation of working solution
Dilute to 1-100 μM with PBS or serum-free cell culture medium (optimized according to the experiment).
Note: The working solution should be prepared and used immediately, and should be protected from light.

2. Cell staining
2.1 Disperse cells with 100 μL PBS and suspend them in a 96-well plate at a density of 2×10⁵.
2.2 Add AkaLumine hydrochloride (100 µM) and 5 mM ATP magnesium (HY-B2176) to the cells.
2.3 Analyze using a bioluminescence imaging system.
In Vivo:Subcutaneous/orthotopic tumor mouse imaging^[1]
1. Inject tumor cells into mice subcutaneously or intratibially.
2. Inject different concentrations of AkaLumine hydrochloride (0.5-33 mM; 100 μL) intraperitoneally.
3. Detect using a bioluminescence instrument 15 minutes after injection.