D-Luciferin, Potassium Salt -40902ES
D-Luciferin, Potassium Salt -40902ES
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D-Luciferin, Potassium Salt -40902ESD-Luciferin, Potassium Salt -40902ES
D-Luciferin, Potassium Salt -40902ES
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 (substrate) can be oxidized to emit light in the presence of ATP and luciferase. When fluorescein is in excess, the quantum number of light produced is equal to Luciferase concentrations were positively correlated (see figure below). Plasmids carrying luciferase encoding gene (Luc) were transfected into cells and introduced into study animals such as rats and mice.   D-luciferin is also commonly used in in vitro...

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D-Luciferin, Potassium Salt -40902ES

YeasenSKU: 40902ES01
Size: 100 mg
سعر:
$100.00
مخزون:
في الأوراق المالية

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 (substrate) can be oxidized to emit light in the presence of ATP and luciferase. When fluorescein is in excess, the quantum number of light produced is equal to Luciferase concentrations were positively correlated (see figure below). Plasmids carrying luciferase encoding gene (Luc) were transfected into cells and introduced into study animals such as rats and mice.

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 three Product form: D-luciferin (free acid), D-luciferin salt (sodium salt and potassium salt). The main difference lies in solubility: water solubility of the former and solubility of the buffer system. They are weakly soluble except in weak bases such as low concentrations of NaOH and KOH solutions. Soluble in methanol and DMSO; The latter can be easily dissolved in water or buffer, easy to use.

Features

No radiation, almost harmless to living organisms.

Bioluminescence, no excitation light source.

So sensitive, you can detect it in a few hundred cells.

Good penetration, 3-4cm tissue depth can still be detected.

High signal-to-noise ratio, strong fluorescence signal and good anti-interference.

Applications

1. In the tumorigenesis experiment in nude mice, the tumor growth was observed without invasion in real time, without tumor stripping measurement.

2. 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.

3. The localization and distribution of foreign cells in animals were detected.

4. 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.

5. 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 potassium salt; D-Luciferin firefly, potassium salt
CAS NO. 115144-35-9
Formula C₁₁H₇N₂O₃S₂K
Molecular weight 318.42 g/mol
Appearance Light yellow powder
Solubility Soluble in water(60 mg/mL)

Components

Components No. Name 40902ES01 40902ES02 40902ES03 40902ES08
40902 D-Luciferin, Potassium Salt 100mg 500 mg 1g 5g

Figures

Cited from "A Novel Allosteric Inhibitor of Phosphoglycerate Mutase 1 Suppresses Growth and Metastasis of Non-Small-Cell Lung Cancer, Cell Metab. 2019;30(6):1107-1119

Storage

The product is shipped with dry ice and can be stored at -15℃ ~ -25℃ for 1 year.

FAQ

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
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.
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 for in vivo imaging and reporter gene systems. It is detected through a cold luminescence module and does not require excitation light stimulation.
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 the reaction toxicity will be smaller (Fluorescein is a low-molecular-weight organic compound composed of phenylthiazole and thiazole carboxyl group, with low toxicity).
3) Usage effect: There is no significant difference. In in vivo experimental studies, the usage rate of potassium salt is higher.
It has an impact. The purity is above 99%, with a good level of 99%. For 99% pure fluorescein, there is 1% solid impurity. If this 1 gram of impurity is dissolved in 25 ml of buffer solution (this dilution ratio is the standard dilution method for in vivo imaging experiments), the concentration of the impurity at this time is 0.4 g/L. Assuming the molecular weight of the impurity is 1000 g/mol, then the molar concentration of the impurity 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.
The powder should be stored in a dark place at -20 or -70 degrees Celsius. Its validity period is at least one year.
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 coelenterazine is an action substrate for various luciferases such as sea kidney luciferase (Rluc) and Gaussia luciferase (Gluc).
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, 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).
Fluorescein is a small molecule with excellent water solubility and lipid solubility. It can easily penetrate the cell membrane and the blood-brain barrier.
Either one is fine. Intraperitoneal injection has a slower diffusion rate and the light emission lasts longer; intravenous injection has a faster diffusion rate, but the light emission lasts for a very short period of time.
Intraperitoneal injection (i.p.), with a fluorescein concentration of 150 mg/kg per kg of body weight. Taking a 20g C57 mouse as an example, the dosage for each mouse for one test: 150 mg/kg × 20g = 3mg, 100mg ÷ 3mg = 33 mice. This is for reference only. The specific dosage should be based on the reagent and without any loss.
Generally speaking, potassium d-fluorescein does not cause any toxic effects on animals.
There is no such information.
This product undergoes in vitro imaging. The ATP and magnesium ions required for this process need to be achieved through the cell culture medium. Therefore, a cell culture medium containing ATP and magnesium ions needs to be added to dilute the probe;
2. The cells themselves also contain a certain amount of ATP and magnesium, which can promote the reaction. In most cases, the ATP and magnesium contained in the cells are sufficient for the reaction of D-fluorescein salt, but the content varies depending on the type of cells. You can first try using a culture medium without additional ATP and magnesium, and conduct the experiment normally to see the results.
Chemiluminescence, no excitation wavelength required. The detection wavelength for firefly luciferin is 560 nm. The detection wavelength for sea kidney luciferin is 465 nm.
It can pass through the blood-brain barrier, the placental barrier and the blood test barrier, and also enter living cells.
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 is: 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.
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.
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.

Citations

Documents

Brochure
In_vivo_imaging_Leaving_Tumor_Cells_Nowhere_to_Hide.pdf

Applications

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