D-Luciferin Potassium Salt

Catalog No.S6580 Batch:S658001

Technical Data

Formula	C₁₁H₇KN₂O₃S₂
Molecular Weight	318.41	CAS No.	115144-35-9
Solubility (25°C)*	In vitro	Water	16 mg/mL (50.24 mM)
		DMSO	4 mg/mL (12.56 mM)
		Ethanol	Insoluble
* <1 mg/ml means slightly soluble or insoluble. * Please note that Selleck tests the solubility of all compounds in-house, and the actual solubility may differ slightly from published values. This is normal and is due to slight batch-to-batch variations. * Room temperature shipping (Stability testing shows this product can be shipped without any cooling measures.)

Preparing Stock Solutions

Biological Activity

Description	D-luciferin is the natural substrate of the enzyme luciferase (Luc) which catalyzes the production of the typical yellow-green light of fireflies. The 560 nm chemiluminescence from this reaction peaks within seconds, with light output that is proportional to luciferase concentration when the substrate luciferin is present in excess. The luciferase (luc) gene is a popular reporter gene for research and agent screening. Chemiluminescent techniques are virtually background-free, making the luc reporter gene ideal for detecting low-level gene expression. In addition to its role as a reporter of gene expression, luciferase is commonly used in an extremely sensitive assay for ATP.
In vitro	1.Example protocol for in vitro bioluminescent image assays a) Prepare a 100 mM (100-200X) Luciferin stock solution in sterile water. Mix well. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light. b) Prepare a 0.5-1 mM working solution of D-Luciferin in pre-warmed tissue culture medium. c) Aspirate media from cultured cells. d) Add Luciferin working solution to cells, and incubate the cells for 5-10 minutes at 37℃ just prior to imaging. 2.Example protocol for in vivo bioluminescent image assays a) Prepare a 15 mg/mL Luciferin stock solution in DPBS, without Mg2+ and Ca2+. Mix well. b) Filter sterilizes the solution through a 0.2 μm filter. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light. c) Inject the luciferin intra-peritoneally (i.p.) 10-15 minutes before imaging at 150 mg/kg (or 10 μL/g of luciferin stock solution) of the animal body weight. Note: A kinetic study of luciferin should be performed for each animal model to determine peak signal time. 3.Example protocol for luciferin reporter assays a) Prepare a 100 mM Luciferin stock solution in sterile water. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light. b) Prepare a 1 mM working solution of D-Luciferin with 3 mM ATP, 1 mM DTT and 15 mM MgSO4 in 25 mM tricine buffer pH 7.8. c) Pipette 5-10 μLof cell lysate into a microplate. Use lysis reagent or buffer without lysate as a blank. d) Prime luminometer with luciferin working solution. e) Inject 200 μL of luciferin working solution with no delay and a 10 second integration time.

Description

D-luciferin is the natural substrate of the enzyme luciferase (Luc) which catalyzes the production of the typical yellow-green light of fireflies. The 560 nm chemiluminescence from this reaction peaks within seconds, with light output that is proportional to luciferase concentration when the substrate luciferin is present in excess. The luciferase (luc) gene is a popular reporter gene for research and agent screening. Chemiluminescent techniques are virtually background-free, making the luc reporter gene ideal for detecting low-level gene expression. In addition to its role as a reporter of gene expression, luciferase is commonly used in an extremely sensitive assay for ATP.

In vitro

1.Example protocol for in vitro bioluminescent image assays
a) Prepare a 100 mM (100-200X) Luciferin stock solution in sterile water. Mix well. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light.
b) Prepare a 0.5-1 mM working solution of D-Luciferin in pre-warmed tissue culture medium.
c) Aspirate media from cultured cells.
d) Add Luciferin working solution to cells, and incubate the cells for 5-10 minutes at 37℃ just prior to imaging.
2.Example protocol for in vivo bioluminescent image assays
a) Prepare a 15 mg/mL Luciferin stock solution in DPBS, without Mg2+ and Ca2+. Mix well.
b) Filter sterilizes the solution through a 0.2 μm filter. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light.
c) Inject the luciferin intra-peritoneally (i.p.) 10-15 minutes before imaging at 150 mg/kg (or 10 μL/g of luciferin stock solution) of the animal body weight.
Note: A kinetic study of luciferin should be performed for each animal model to determine peak signal time.
3.Example protocol for luciferin reporter assays
a) Prepare a 100 mM Luciferin stock solution in sterile water. Use immediately, or make single use aliquots, and store at -20℃, avoid freeze-thaw cycles, avoid exposure to the light.
b) Prepare a 1 mM working solution of D-Luciferin with 3 mM ATP, 1 mM DTT and 15 mM MgSO4 in 25 mM tricine buffer pH 7.8.
c) Pipette 5-10 μLof cell lysate into a microplate. Use lysis reagent or buffer without lysate as a blank.
d) Prime luminometer with luciferin working solution.
e) Inject 200 μL of luciferin working solution with no delay and a 10 second integration time.

Protocol (from reference)

Animal Study:^{^[2]}	Animal Models Female NCI Athymic nu/nu nude mice Dosages 3 mg/100 μL Administration i.p.

References

Selleck's D-Luciferin Potassium Salt has been cited by 4 publications

Carbenoxolone disodium suppresses the migration of gastric cancer by targeting HDAC6 [ Future Med Chem, 2023, 15(4):333-344]	PubMed: 36946221
A magneto-controlled biocatalytic cascade with a fluorescent output [ Org Biomol Chem, 2022, 20(9):1869-1873]	PubMed: 35156979
Leronlimab, a humanized monoclonal antibody to CCR5, blocks breast cancer cellular metastasis and enhances cell death induced by DNA damaging chemotherapy [ Breast Cancer Res, 2021, 23(1):11]	PubMed: 33485378
KDM4A-mediated histone demethylation of SLC7A11 inhibits cell ferroptosis in osteosarcoma [ Biochem Biophys Res Commun, 2021, 550:77-83]	PubMed: 33689883

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SHIPPING AND STORAGE
Selleck products are transported at room temperature. If you receive the product at room temperature, please rest assured, the Selleck Quality Inspection Department has conducted experiments to verify that the normal temperature placement of one month will not affect the biological activity of powder products. After collecting, please store the product according to the requirements described in the datasheet. Most Selleck products are stable under the recommended conditions.

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