1. What experiments are suitable for D-fluorescein?
D-fluorescein, specifically D-luciferin potassium salt and 5-fluoro-luciferin, serves as a key substrate for luciferase in various experimental applications. These compounds are widely used in: - **Reporter gene assays** to monitor gene expression - **In vivo imaging** to visualize biological processes in living organisms - **In vitro imaging** for cell-based studies and real-time monitoring - **ATP quantification** due to the direct correlation between ATP levels and luminescence intensity - **Luciferase activity analysis** and related genetic research projects These applications make D-fluorescein an essential tool in molecular biology, pharmacology, and biomedical research.2. The basic principle of D-fluorescein in in vivo imaging
The luminescent reaction involving D-luciferin and luciferase follows a standard enzymatic mechanism. When D-luciferin is present in excess, the amount of light emitted (photons) is directly proportional to the number of luciferase molecules and the number of cells. This enables researchers to quantify cell populations or track gene expression over time in live animals, making it a powerful tool for non-invasive imaging.3. Stability of D-fluorescein potassium salt solution
D-luciferin potassium salt dissolves easily in water and buffer solutions, with a solubility of up to 25 mg/mL. It is typically used at concentrations ranging from 3 to 15 mg/mL. However, its stability is influenced by several factors: - **pH**: Solutions with a pH below 6.5 may lead to hydrolysis, while a pH above 7.5 can cause racemization, converting D-luciferin into the L-form, which is less effective. - **Oxygen exposure**: Even small amounts of oxygen can accelerate degradation. - **Storage conditions**: Under optimal storage (-80°C, low oxygen, and appropriate pH), D-luciferin degrades at a rate of 0.2% per day. Therefore, it should be used within one year after preparation. Additionally, D-luciferin tends to racemize in the presence of water, reducing its effectiveness in luciferase reactions.4. Differences between D-luciferin, sodium D-luciferin, and potassium D-fluorescein
The primary distinction among these forms lies in their solubility and dispersion properties. D-luciferin potassium salt has superior solubility in aqueous solutions, making it ideal for most biological assays. Sodium D-luciferin may have similar performance but is less commonly used due to lower solubility. Potassium D-fluorescein is preferred in many laboratory settings because of its fast dissolution and high compatibility with biological systems.5. Storage and transportation conditions for D-fluorescein potassium salt
To maintain its stability, D-luciferin potassium salt should be stored at ≤ -20°C, away from light, and in a dry environment. The unopened bottle is typically filled with argon gas to prevent oxidation and degradation. It can remain stable for up to two years if unopened. During transportation, high-purity D-luciferin potassium salt can be kept at room temperature for short periods. However, once the package is opened, strict storage conditions must be followed. When weighing D-luciferin potassium salt, it’s important to note that it is hygroscopic. To avoid moisture absorption, the sample should be allowed to reach room temperature before weighing, and the process should be completed quickly. Proper handling ensures consistent performance in experiments.latex dog toy,durable latex dog toys,squeaky latex dog toys,latex dog squeaky toy,small latex dog toys,Squeaky Dog Latex Toy,Sound Pet Toy Crocodile,Dog Latex Toy Crocodile,Latex Squeaky Dog Chew Toy
Dongguan King Pet Toys Co.,Ltd , https://www.kingpettoys.com