Luciferase from Photinus pyralis (firefly)
SIGMA/SRE0045 - recombinant, expressed in E. coli, lyophilized powder, ≥10×1010 units/mg protein
Synonym: Luciferase firefly
CAS Number: 61970-00-1
MDL Number: MFCD00131546
Product Type: Chemical
ATP is hydrolyzed and light is emitted when firefly luciferase catalyzes the oxidation of D-luciferin. Reaction (1) is reversible and the equilibrium lies far to the right. Reaction (2) is essentially irreversible. When ATP is the limiting reagent, the light emitted is proportional to the ATP present.
10% SDS-PAGE analysis of recombinant luciferase SRE0045
An ATP dose response curve was performed in 50 μL reaction cocktails containing, decreasing amounts of ATP, 0.2 μg Luciferase (Cat. No. SRE0045), and 15 nmol luciferin in a Tris-glycine buffer, pH 7.6 supplemented with 0.5 mM EDTA, 5 mM MgSO4 and 0.1% BSA. Luminescence was measured at 22 °C for 10 seconds in quadruplicates, using a GloMax 20/20 Luminometer.
| application(s) | diagnostic assay manufacturing |
| form | lyophilized powder |
| mol wt | 62 kDa |
| Quality Level | 200  |
| recombinant | expressed in E. coli |
| specific activity | ≥10×1010 units/mg protein |
| storage temp. | −20°C |
| Application: | Firefly luciferase is used extensively in molecular and cell biology, in particular for the efficient detection and quantitation of ATP and as a reporter for genetic function. |
| Biochem/physiol Actions: | Firefly luciferase is a 62 kDa protein that catalyzes the production of light. The enzyme requires ATP, molecular oxygen, and the heterocyclic compound, firefly luciferin, to generate light in a two-step process. The light producing reaction is initiated by luciferin activation (adenylation of its carboxylate group) and proceeds in the presence of molecular oxygen to yield a photon of yellow-green light. |
| Other Notes: | One luciferase enzyme unit will produce one Relative Light Unit (RLU) at 20-25 °C over a 10 second period, measured in 100 μl assay mixture containing 40 pmol ATP and 15 nmol luciferin in Tris-glycine buffer, pH 7.6, using a GloMax 20/20 Luminometer. Unit Definition Conversion Factor: There are approximately 9000 Relative Light Units (RLU) per one traditional Light Unit that uses a peak height equivalent to 0.02 μCi of 14C in a PPO/POPOP cocktail. |
| Physical form: | Supplied as a lyophilized powder containing HEPES, pH 7.5, NaCl, MgCl2, EDTA, DTT and a carbohydrate stabilizer. |
| Physical form: | This product is a recombinant luciferase (62 kDa) from Photinus pyralis (American firefly) produced from the luc gene expressed in E. coli. |
| Preparation Note: | To obtain maximal solubility it is important to reconstitute the enzyme at a high salt concentration, such as 1 M Tris buffer with any counter ion at pH 7-8. The enzyme can be prepared at a concentration of up to 5 mg protein/ml. Do not vortex and avoid agitation. After reconstitution, the enzyme solutions can kept at 4-8 °C for up to 2 days or frozen in working aliquots at -20°C for at least one month. Repeated freezing and thawing is not recommended. |
| Symbol |  GHS05 |
| Signal word | Danger |
| Hazard statements | H318 |
| Precautionary statements | P280 - P305 + P351 + P338 |
| RIDADR | NONH for all modes of transport |
| WGK Germany | WGK 3 |
| Flash Point(F) | Not applicable |
| Flash Point(C) | Not applicable |
| activity | specific activity: ≥10×1010 units/mg protein |
| Storage Temp. | −20°C |
| Enzyme Commission (EC) Number | 1.13.12.7 ( BRENDA  | IUBMB ) |
| UNSPSC | 12352204 |