Product Description
D-Luciferin is the most popular and multifunctional bioluminescent substrate. The luciferase/luciferin bioluminescence system of fireflies is present in fireflies (Photonus pyralis) and several other beetles. Luciferase oxidizes ATP activated luciferin through the intermediate of dioxanone. Firefly luciferase produces light through ATP dependent oxidation of luciferin. The 560nm chemiluminescence from this reaction reaches its peak within seconds, and when there is an excess of luciferin and ATP, the light output is proportional to the activity of luciferase. Firefly luciferase has long been bound to antibodies and used as a marker in immunoassays using luciferin as a detection substrate. Compared with HRP and alkaline phosphatase, luciferase has poorer tolerance to chemical modifications. A special advantage of this enzyme is that, in addition to its high sensitivity, it has low endogenous luciferase activity in mammalian tissues. Another important use of luciferase is in the field of health monitoring. The luciferase/luciferin system can be used to detect pollution, as ATP present in all organisms needs to produce luminescence. The main application of this ATP bioluminescence is to determine quality by testing the surface in food processing plants to determine the presence of equipment or product contamination.
D-luciferin is also commonly used for in vitro research, including analysis of luciferase and ATP levels; Reporting gene analysis; High throughput sequencing and various pollution detection. There are currently three product forms on the market, D-fluorescein (free acid), D-fluorescein sodium salt, and D-fluorescein potassium salt. The main difference between these three products lies in their dissolution characteristics. D-fluorescein (free acid) has weak water solubility and buffering system solubility, unless dissolved in weak alkaline solutions such as NaOH and KOH. Dissolved in methanol (10 mg/mL) and DMSO (50 mg/mL). However, D-fluorescein in the form of sodium and potassium salts can be easily and quickly dissolved in water or buffer solutions, making it convenient to use and the solvent non-toxic, making it particularly suitable for in vivo experiments. There is no substantial difference in the vast majority of applications among these three products that have been formulated into liquids.
ordering information
Product Name | Article number | Specifications |
D-Fluorescein Sodium Salt (Ultra Pure)(D-Luciferin, sodium salt) | AC19L111 | 25 mg |
D-Fluorescein Sodium Salt (Ultra Pure)(D-Luciferin, sodium salt) | AC19L112 | 100 mg |
D-Fluorescein Sodium Salt (Ultra Pure)(D-Luciferin, sodium salt) | AC19L113 | 1 g |
Product nature
Chinese alias（Chinese synonym） | D-Fluorescein Sodium Salt |
English alias（English synonym） | D-Luciferin, sodium salt |
CAS number（CAS NO.） | 103404-75-7 |
molecular weight（Molecular weight） | 302.3 g/mol |
Solubility（Solubility） | dissolve in water |
Transportation and Preservation
Blue ice transportation, stored at -20 ℃. The validity period is 24 months.
usage method
1. Implementation plan for in vitro bioluminescence image measurement
（1）Prepare 100mM (100-200X) fluorescent solution in sterile water. Mix evenly. Immediately use, or use equal parts of the sample separately, and store at -20 ℃ to avoid freeze-thaw cycles and exposure to light.
（2）Prepare a working solution of 0.5-1mM D-luciferin in preheated tissue culture medium.
（3）Separate the culture medium from the cultured cells.
（4）Add the working solution of fluorescein to the cells and incubate the cells at 37 ℃ for 5-10 minutes before imaging.
2. Implementation plan for in vivo bioluminescence image measurement
（1）Prepare a 15mg/mL fluorescent reserve solution in DPBS, free of Mg2+and Ca2+. Mix evenly.
（2）Filter the sterilization solution through a 0.2 μ m filter. Immediately use, or use equal parts of the sample separately, and store at -20 ℃ to avoid freeze-thaw cycles and exposure to light.
（3）Inject fluorescein into the peritoneum (i.p.) 10-15 minutes before imaging with an animal weight of 150mg/kg (or 10 μ L/g of fluorescein reserve solution).
[Note]: A kinetic study of fluorescein should be conducted on each animal model to determine the peak signal time.
3. Implementation plan for fluorescence report analysis and molecular determination
（1）Prepare a 100mM fluorescent reserve solution in sterile water. Immediately use, or use equal parts of the sample separately, and store at -20 ℃ to avoid freeze-thaw cycles and exposure to light.
（2）Prepare a working solution of 1mM D-fluorescein, containing a 25mM tricine buffer containing 3mM ATP, 1mM DTT, and 15mM MgSO4 at pH 7.8.
（3）Transfer 5-10 μ L of cell lysate into a microplate. Use a lysis reagent or buffer without lysis solution as a blank.
（4）According to the manufacturer's instructions, use a Puli spectrophotometer with a fluorescent working solution.
（5）Inject 200 μ L of fluorescent working solution without delay, with a 10 second integration time.
matters needing attention
1. This product is limited to scientific experimental research and cannot be used in clinical diagnosis, treatment, or other fields.
2. D-fluorescein potassium salt is soluble in sterile water and buffer solution, with a solubility of up to 25mg/mL. The general usage concentration is 3-15 mg/mL. The pH value of the solution, oxygen in the solution, and storage time are very important for its stability during storage. When the pH of the solution (undergoes hydrolysis) or>7.5 (undergoes racemization, converting D-type to L-type), D-fluorescein potassium salt is relatively unstable. If there is a small amount of oxygen in the solution, it will accelerate the degradation rate of D-fluorescein potassium. The reserve solution can be prepared in ATP free water and stored in a dark place at -20 ℃. Free acids must be neutralized with appropriate alkalis for dissolution.
3. D-fluorescein can be used in conjunction with any existing literature or ATP analysis system.
4. If detecting ATP, please wear gloves and use an ATP free container to minimize all possible sources of ATP contamination. Only use sterile ATP free water and reagents. Prepare all reagents using high-pressure sterilized water.
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This product is limited to scientific experimental research and cannot be used in clinical diagnosis, treatment, or other fields.
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