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Cat. No.: DLP-100 (for 100mg)

Cat. No.: DLP-1k (for 1g)

Description

D-Luciferin Potassium (CAS: 115144-35-9) is a high-signal, high-stability substrate for detecting firefly luciferase activity. Once prepared as a liquid, this product should be stored at -80°C; its effectiveness may gradually decline if stored at -20°C for extended periods. The performance of this product meets or even exceeds that of leading similar products in certain aspects.

Advantage

This product offers high luminescence intensity with stable signal performance. It provides strong luminescence, suitable for various routine assays. The signal stability is relatively good: within 1 minute, the signal remains stable with fluctuations not exceeding 8%; within 3 minutes, fluctuations do not exceed 20%; and within 5 minutes, fluctuations are under 35%. The signal half-life is approximately 8 minutes, making it particularly advantageous for assays with a large number of samples, such as in 96-well plates for firefly luciferase activity assays.

Principle

Firefly luciferase is a protein with a molecular weight of about 61 kD that catalyzes the oxidation of luciferin to oxyluciferin in the presence of ATP, magnesium ions, and oxygen. This oxidation process produces bioluminescence, which can be measured using a luminometer or a liquid scintillation counter.

The bioluminescence system using luciferin and luciferase allows for highly sensitive and efficient detection of gene expression. Typically, regulatory elements or the 5' promoter region of the gene of interest are cloned upstream of luciferase, or the 3'-UTR is cloned downstream of luciferase to construct a reporter gene plasmid. The plasmid is then transfected into cells, which are subsequently treated with appropriate drugs and lysed. Luciferase activity is measured to assess the impact of drug treatments on the transcriptional regulation of the target gene.

Storage

Store at -20°C in the dark; remains effective for at least two years.

Note

• To achieve the best measurement results, when using a single-tube chemiluminescence instrument, try to keep the time between mixing the sample and the assay reagent and the measurement as consistent as possible, for example, within 30 seconds. When using a multifunctional fluorescence microplate reader with chemiluminescence measurement capabilities, it is advisable to add all the samples first and then uniformly add the D-Luciferin potassium.
• Since luciferase activity is sensitive to temperature, both the sample and the assay reagent should be at room temperature before measurement. The firefly luciferase detection buffer can be thawed and mixed at room temperature or in a water bath not exceeding 25°C before being mixed with the assay substrate.
• To ensure the stability of the luciferase assay and achieve optimal results, it is recommended to prepare the assay reagents fresh for each use. Unused reagents can be aliquoted and stored in the dark to avoid repeated freeze-thaw cycles and prolonged exposure to room temperature.
• Use white or black 96-well plates for detection. Ordinary transparent 96-well plates may cause interference between adjacent wells.
• After mixing the sample with the assay reagent, wait for 1-2 seconds before measurement. The measurement time is typically 10 seconds, but can be adjusted based on the situation. However, for consistency, use the same measurement time for samples from the same batch.
• To avoid errors due to differences in transfection efficiency, you can co-transfect a reporter gene plasmid for Renilla luciferase as an internal control, or co-transfect a reporter gene plasmid for β-galactosidase (β-gal) as an internal control.
• This product is intended for scientific research by professionals only and must not be used for clinical diagnosis or treatment, food, or pharmaceuticals. It should not be stored in ordinary residential areas.
• For your safety and health, please wear laboratory coats and disposable gloves during handling.

Only for research and not intended for treatment of humans or animals

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