AAT Bioquest

How to prepare Luciferin for in vivo use?

Posted August 5, 2019

Luciferin is a common reagent used for in vivo imaging of luciferase labeled tumors or other infectious diseases in rats or mice. Below is an example protocol for in vivo bioluminescent imaging using D-Luciferin, free acid. 
  • D-Luciferin, free acid (Cat No. 12501, 12502 and 12503)
  • 1X dPBS w/o Ca2+ and Mg2+
  • Syringe filter, 0.22 µm
  1. Prepare a fresh stock solution of D-Luciferin, free acid at 15 mg/mL in 1X dPBS w/o Ca2+ and Mg2+.
  2. Filter through a 0.22 µm syringe filter to sterilize solution.
  3. Use immediately, or aliquot into sterilized tubes and freeze at -80°C for up to 6 months.
  4. Inject the Luciferin solution intra-peritoneally 10-15 minutes before imaging*.
  5. Inject 10 µL/g of body weight. Each mouse should receive 150 mg Luciferin/kg of body weight.
  6. Take images every 5-10 minutes for about 40-60 minutes to determine the kinetic curve.
*A kinetic study of luciferin should be performed for each animal model to determine peak signal time.
Additional resources

D-Luciferin Product Information Sheet