Content and storage

General Specifications 

Color:  Light yellow powder 

Detection Method: Bioluminescence

Excitation Class: Visible 

Molecular Weight: 280.32 g.mol^-1

Formula: C₁₁H₈N₂O₃S₂

Cas Number: 2591-17-5 

Shipping Condition: Dry Ice 

Regulatory Statement: For Research Use Only. Not for use in diagnostic procedures. 

Description

Luciferin is a small molecule that consists of a benzothiazole moiety attached to a thiazole carboxylic acid moiety. Luciferin is a natural substrate of luciferase, an enzyme found in Firefly luciferase. This molecule has fluorescent properties (Ex 328 nm; Em 532 nm in H₂O)^1,2 but it is mainly used for bioluminescent imaging (BLI) purpose.

Bioluminescence is a natural process that has been found in various living organisms such as the North American firefly (Photinus pyralis) and is based on the oxidation of D-luciferin catalyzed by the enzyme Luciferase. Upon recognition by the enzyme, D-luciferin is oxidized to oxy-luciferin and releasing one photon of light, which can be detected by a CCD camera (Figure 1). The intensity of the light output is closely related to the amount of D-luciferin available for the enzyme and therefore it is possible to quantify the amount of luciferin by measuring the amount of emitted light with a CCD camera. As a non-invasive imaging method, BLI is comparable to other in vitro and in vivo techniques but has the advantage of high sensitivity, convenience and ease of use. It does not require a light source (as opposed to fluorescence) and there is no background signal from tissues that do not express luciferase. Moreover, it allows real-time imaging of luciferase expressing cells or luciferase expressing mice.^3-5 The luciferin/Luciferase process is very substrate dependent and does not allow significant modification on the luciferin scaffold to be recognized by the enzyme.

Figure 1: Oxydation of D-luciferin by Firefly luciferase forming oxy-luciferin and a photon of light

Guideline for use

D-luciferin for in vitro use:

– 1g of D-luciferin is dissolved in 33.3 mL of sterile water to make a 30 mg/mL stock solution. After mixing and filtering (0.2 um), the solution is aliquoted and purged with nitrogen (inert gas prevents oxidation) protect from light and freeze down to -80°C for future use. Frozen stock solutions can be stored up to 1 year.

– D-luciferin stock solution is thawed, kept on ice and protected from light. Stock solution is diluted at 1:200 in complete culture medium to 150 ug/mL. Luciferase expressing cells are incubated 5 min at 37°C prior to imaging. Diluted solution should be discarded after use.

D-luciferin for in vivo use:

– 1g of D-luciferin is dissolved in 66.6 mL of DPBS, w/o Mg²⁺ and Ca²⁺ to make a 15mg/mL solution. After mixing and filtering, the solution is aliquoted and purged with nitrogen (inert gas prevents oxidation) protect from light and freeze down to -80°C for future use. Frozen stock solution can be stored up to 1 year.

– D-luciferin solution is thawed, kept on ice and protected from light. Diluted solution should be discarded after use

– Mouse is injected IP with D-luciferin solution at 150mg/kg of body weight. Images are acquired after 10-15 min post-injection.

References

1. White, E.H. et al.,(1963) J. Am. Chem. Soc., 85, 337

2. Bowie, L.J., (1978) Methods in Enzymology, 57, 23

3. Prescher, J. A., and Contag, C. H. (2010) Curr. Opin. Chem. Biol. 14, 80-89.

4. McCaffrey, A., Kay, M. A., and Contag, C. H. (2003) Mol. Imaging 2, 75-86

5. Massoud, T. F., and Gambhir, S. S. (2003) Genes Dev. 17, 545-580.

Price

Code No.	Product	Size	Price	Protocol	MSDS
RS101-01	D-Luciferin potassium salt (Standard Purity)	1g	$680	–	–

Contact Information

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