高穩定性StayBrite™ D-Luciferin, Sodium Salt
Catalog#: 7902 -1G | Size: 1 g
Alternate Name/Synonyms: D-Luciferin, Sodium Salt, StayBrite™
Appearance: Light yellow solid
Formulation: N/A
CAS Number: 103404-75-7
Molecular Formula: C₁₁H₇N₂O₃S₂. Na
Molecular Weight分子量: 302.3
Purity純度: ≥98% by HPLC
Solubility: H₂O
Storage Temp.: -20°C
Shipping Conditions: gel pack
Description: A useful substrate for the enzyme Luciferase. Upon oxidation by the enzyme luciferase, produces bioluminescence.
Handling: Protect from air and light
7902-100 | StayBrite™ D-Luciferin, Sodium Salt | 100 mg |
7902 -1G | StayBrite™ D-Luciferin, Sodium Salt | 1 g |
7902 -10G | StayBrite™ D-Luciferin, Sodium Salt | 10 g |
Bacterial luciferase uses the substrate bacterial luciferin which is a different molecule from D-luciferin. The plasmid encoding for bacterial luciferase also encodes for the substrate which is then synthesized by the cell, eliminating the neeed for exogenous substrate administration.
Renilla and Gaussia luciferase uses the substrate coelenterazine, again different from D-luciferin.
Firefly or clickbeetle luciferase catalyzes the oxidization of D-luciferin to oxyluciferin in the presence of ATP, Mg2+, and oxygen with production of a yellow-green light, shifting to red light in vivo at 37oC. Since ATP is required as a co-factor, Firefly luciferase can be used as an indicator of the presence of energy or “life” and function as a life-death stain.
Methods:
Dissolution
D-luciferin for in vitro use:
Dissolve 1g of D-luciferin in 33.3ml sterile water to make a 30 mg/ml * stock solution.
Mix gently, syringe filter (0.2 um), aliquot, purge with nitrogen or argon (inert gas prevents oxidation) and freeze under at -80°C for future use (up to 1 year), protect from light.
Thaw and keep on ice, protect from light. If the luciferin does not dissolve, briefly bring to temperature in a waterbath and gently mix. Dilute stock at 1:200 in complete tissue culture medium to 150ug/ml . incubate 5 min prior to imaging.
Dispose at end of day.
D-luciferin for in vivo use:
Dissolve 1g of D-luciferin in 66.6 ml in DPBS, w/o Mg2+ and Ca2+ to make a 15mg/ml solution*. Mix gently, syringe filter (0.2 um), aliquot, purge with nitrogen or argon (inert gas prevents oxidation) and freeze at -80°C for future use (up to 1 year), protect from light.
Thaw and keep on ice, protect from light. If the luciferin does not dissolve, briefly bring to temperature in a waterbath and gently mix. Dispose at end of day.
Inject at 150mg/kg, thus 200ul (3mg) for a 20 g mouse. Image in plateau phase (see kinetics curve).
1 g is sufficient for about 300 mice.
* D-luciferin can be dissolved up to a concentration of 50mg/ml or 178mM, it precipitates out at 60mg/ml. Na+ salt dissolves better than K+ salt, to 100mg/ml.
Injection routes:
Intraperitoneal (IP) injection:
injection of the substrate in the intraperitoneal cavity. A dose of 150mg/kg is recommended. Inject 200ul (of a 15mg/ml solution) for a 20 g mouse. (The maximum IP volume for a 20 g mouse is 1ml). Use a 25 G needle, 1 cc syringe. Take mouse in one hand, restrain the mouse belly up, hold the scruff of the neck between thumb and index and clamp the tail with the other fingers, point the head down (intestinal organs sag cranially). Inject half the dose in the lower left quadrant and the other half in the lower right quadrant for the most consistent results. Insert needle 5mm, bevel side up at a 30o angle.
Intravenous (IV) injection:
Intravenous injections in mice are routinely performed in the tailvein or retroorbital plexus. Even though an intravenous injection is a bolus injection, the same dose of 150mg/kg is recommended. Keep in mind that the maximal volume to inject IV is 10% of the blood volume. A 20g mouse is estimated to have 2ml blood, thus the maximal IV injection volume is 200ul. Use a 1 cc syringe and 26G needle or an insulin syringe with 28G needle. Make sure no airbubbles are present in the syringe. Place the animals under a heating lamp for about 5 minutes or soak the tail in warm water to dilate the tail veins. Put the mouse in a restrainer. Wipe tail with ethanol. Inject the dose slowly
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