D-Luciferin, Sodium Salt D _ 40901ES
D-luciferin is a common substrate for Luciferase and is widely used throughout biotechnology, especially in vivo imaging technology. The mechanism of action is that luciferin (the substrate) is oxidized to emit light in response to ATP and luciferase (see figure below). It's produced when there's too much luciferin The light quantum number was positively correlated with the concentration of luciferase. Plasmids carrying luciferase encoding gene (Luc) were transfected into cells and introduced into study animals such as rats and mice In vivo, fluorescein is then injected and changes in light intensity are detected using bioluminescence imaging (BLI) to monitor disease progression or drug efficacy in real time and so on. ATP's influence on the reaction system can also be used to indicate energy or vital signs according to changes in bioluminescence intensity. D-luciferin is also commonly used in in vitro studies, including luciferase and ATP levels analysis; Reporter gene analysis; High-throughput sequencing and various contamination tests. There are currently three product forms: D-luciferin (free acid), D-luciferin salts (sodium and potassium salts). The main difference lies in solubility: the former is water-soluble and the solubility of buffer system is weak, except soluble in weak bases such as low concentration NaOH and KOH solutions, soluble in methanol and DMSO; The latter is easily soluble in water or buffer, easy to use, solvent non-toxic, especially suitable for in vivo experiments. As a solution, there is no substantial difference between the three products in the vast majority of applications. Features High Purity: >99% High Solubility: Up to 60 mg/mL in water High Sensitivity: Capable of detecting as few as hundreds of cells; effective imaging depth of 3–4 cm Stable Signal: High bioluminescence intensity and slow signal decay, ideal for large-scale in vivo validation studies Application In the tumorigenesis experiment in nude mice, the tumor growth was observed without invasion in real time, without tumor stripping measurement. To test the effect of the administration on tumor growth or metastasis, the fluorescein substrate can be completely eliminated within 3 hours, without interference to the drug experiment. The localization and distribution of foreign cells in animals were detected. The target gene or promoter of the target gene is fused to the luciferase gene to detect changes in gene expression during drug treatment or disease progression. Monitoring stem cell transplantation, survival and proliferation; Trace the distribution and migration of stem cells in vivo. Specifications English synonym (S)-4,5-Dihydro-2-(6-hydroxy-2-benzothiazolyl)-4-thiazolecarboxylic acid sodium salt; D-Luciferin firefly, sodium salt monohydrate; CAS NO. 103404-75-7 Formula NaC11H7N2O3S2·H2O Molecular weight 320.32 g/mol Appearance Light yellow powder Solubility Solube in water(100 mg/mL) Purity (HPLC) ≥95% Components Components No. Name 40901ES01 40901ES02 40901ES03 40901ES08 40901ES10 40901 D-Luciferin, Sodium Salt 100mg 500 mg 1g 5g 10g Storage The product should be stored at -15℃ ~ -25℃ for 1 year. Figures 1. Superior Performance (A). Superior Signal Stability Figure 23. Superior Signal Stability Over Time Fluorescence signal was measured every 5 min. Results showed that the signal from the Yeasen product decayed more slowly, with only about 20% reduction (B). Higher Fluorescence Intensity Figure 24. Higher Fluorescence Intensity Fluorescence intensity test using a multifunctional microplate reader demonstrated that the Yeasen luciferin potassium salt produced stronger fluorescence signals compareafter 40 minutes. 1. Application Case Cas 1. D-Luciferin is applicable in cancer research. Figure 25. Mechanistic Study of Circular RNA in Tumor Growth. Using Yeasen products, Gu Y, Wang Y, et al. investigated the role of circRNA in tumor progression. The study revealed that circular RNA circIPO11 drives self-renewal of liver cancer–initiating cells via Hedgehog signaling (Mol Cancer, 2021, 20(1):132). Cas 2. D-Luciferin enables real-time monitoring of tumor growth. Figure 26. In Vivo Pharmacodynamic Evaluation of Antitumor Drugs. Yeasen products were applied in the study by Huang K, Liang Q, et al., which identified a novel allosteric inhibitor of phosphoglycerate mutase 1 (PGAM1) that suppresses growth and metastasis of non–small-cell lung cancer (Cell Metab, 2019, 30(6):1107–1119.e8). FAQ Q: Are there any differences between Firefly Luciferin, Beetle Luciferin and D-Luciferin? A: There is no difference. The three are merely variations in naming by different companies, and they are all compounds (S)-2-(6-Hydroxy-2-benzothiazolyl)-2 - Thiazoline-4-carboxylic acid. Q: What applications do this series of products mainly serve? A: Besides being used for in vivo imaging, fluorescent dye products are also employed in other applications involving luciferase, such as in vitro reporter gene assays, microbial/viral monitoring, pyrosequencing, etc. Q: What is the difference between sodium fluorescein and D-fluorescein sodium? A: Sodium fluorescein: It can be used for cell permeability testing. The permeability of the pigment can be determined by measuring the absorbance value at OD490. D-Fluorescein sodium salt: Used in in vivo imaging and reporter gene systems. It is detected through a cold luminescence module and does not require excitation light stimulation. Q: What are the differences between potassium salt, sodium salt and free acid of fluorescein? The main differences among the three are as follows: 1) Solubility: The salt form is easily soluble in water. The solubility of potassium salt is 60mg/ml, and that of sodium salt is 100mg/ml. Free acid is not easily soluble in water. It can be dissolved by a weak alkaline solution of sodium bicarbonate, and its solubility in methanol is 10mg/ml, and in DMSO is 50mg/ml. 2) Toxicity: The salt form is more convenient to use during the process, especially in in vivo imaging experiments, as it can dissolve in water and thus has less reaction toxicity (Fluorescein is a low-molecular-weight organic compound composed of phenylthiazole and thiazole carboxylic acid groups, which has low toxicity). 3) Usage effect: There is no significant difference. In in vivo experimental studies, the usage rate of potassium salt is higher. Q: Does the purity of the fluorescent dye have any impact on the experiment? A: It has an impact. The purity is above 99%, with a good level of 99%. For 99% pure fluorescein, there is 1% of solid impurities. If these 1 grams of impurities are dissolved in 25 ml of buffer solution (this dilution ratio is the standard dilution method for in vivo imaging experiments), the concentration of the impurities at this time is 0.4 g/L. Assuming the molecular weight of the impurities is 1000 g/mol, then the molar concentration of the impurities is 400 μM. This concentration may inhibit the action of certain enzymes in the cells and may reduce the experimental effect or cause harm to the animals. Q: How stable is the product? A: The powder should be stored in a dark place at -20 or -70 degrees Celsius. Its validity period is at least one year. Q: How does the substrate for in vivo imaging work? A: D-fluorescein (D-Luciferin) is a commonly used substrate for luciferase. Its mechanism of action is that under the action of ATP and luciferase, the fluorescent substance (the substrate) can be oxidized and emit light. When there is an excess of fluorescent substance, the number of photons produced is positively correlated with the concentration of luciferase. 2. Native ctenoidin (Coelenterazine native) is an acting substrate for various luciferases such as sea kidney luciferase (Rluc) and Gaussia luciferase (Gluc). Q: Are there any recommended instruments? Can the multi-functional microplate reader be used? A: Recommended instruments: 1. Instruments with a biochemiluminescence detection module. The light produced by luciferin can be detected by a photometer or a scintillation counter. Common in vivo imaging instruments: such as the IVIS® Lumina small animal in vivo imaging system, and the In-Vivo Xtreme multi-mode small animal in vivo imaging instrument from Bruker, Germany. 2. Multi-functional microplate readers: It is necessary to confirm with the instrument manufacturer whether it is a specific biochemiluminescence detection module (note: cannot use a fluorescence microscope). Q: What are the excitation wavelength and emission wavelength of the substrate? A: Chemiluminescence, no excitation wavelength required. The emission wavelength of firefly luciferin is 560 nm. The emission wavelength of sea kidney luciferin is 465 nm. Q: Can the substrate enter living cells? A: It can pass through the blood-brain barrier, the placental barrier and the blood test barrier, and also enter living cells. Q: Regarding the injection method and dosage of fluorescent substances? A: The recommended concentrations are derived from the literature: 1) Injection method: It can be done through intraperitoneal injection or tail vein injection. 2) Injection volume: The scientific approach is to evaluate the injection dose based on the kinetic curve. The initial attempted injection dose was set at 150mg substrate/kg of mouse body weight. Therefore, the purchase quantity can be calculated according to the above method: If there are 10 mice, 22 - 25g, then 33 - 37.5mg of substrate is required. Q: How does the luminescence property of fluorescein work? A: Approximately 3 minutes after intraperitoneal injection of fluorescein into mice, cells expressing luciferase began to emit light. The intensity reached a stable peak after 10 minutes, and this peak lasted for about 10 - 15 minutes before gradually diminishing. Detection can be conducted within 10 - 15 minutes after the injection. This is for reference only. It is recommended to conduct a preliminary experiment to establish the kinetics curve of luciferase, thereby determining the optimal time for signal detection and the signal plateau period. Q: What are the reasons for the lack of effect in the live imaging experiment? A: To successfully conduct the in vivo imaging experiment, the following conditions are required: The target tissue or cells must express the luciferase gene; the injection of the luciferin substrate must be successful; and the thickness of the tissue at the luminescent site is also a factor. If the experiment fails, the reasons can be investigated from these aspects: whether the luciferase gene is expressed, whether the luciferin substrate was injected correctly, and whether the luminescent site is deep. Documents: Safety Data Sheet 40901_MSDS_HB251208 Manuals 40901_Manual_HB20250520 Related Blog In vivo imaging "detection" technology makes "undercover" cells nowhere to hide Bioluminescence Imaging: A Powerful Tool for Real-Time, In Vivo Research Coelenterazine: one kind of fluorescein reagents widely used in vivo imaging and reporter gene detection Citations & References: [1] Guo Y, Guo Y, Chen C, et al. 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