L9506
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 | SDS-PAGE (4–20%) of luciferase (Cat. No. L9506) under reducing conditions in 1 M Glycine-Tris Buffer with 10 mM Ethylenediaminetetraacetic Acid and 100 mM Magnesium Sulfate, pH 7.0 and stained with Coomassie Blue |
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L9506
Sigma
Luciferase from Photinus pyralis (firefly)
BioUltra, lyophilized powder, ≥98% (SDS-PAGE), 15-30 × 106 light units/mg protein
| Synonym: | Luciferase firefly |
| CAS Number: | 61970-00-1 |
| Enzyme Commission (EC) Number: | 1.13.12.7 ( BRENDA | IUBMB ) |
| MDL number: | MFCD00131546 |
Description
| Analysis Note | Two contaminant, ATP-consuming activities are assayed for in this product, ATPase and nucleoside diphosphokinase. These impurities are found to be less than 5 nanomolar units/mg protein and less than 20 nanomolar units/mg protein, respectively. |
| Note: Prior to 1991, a unit of firefly luciferase activity was defined as that amount which will produce 1.0 nanomole of pyrophosphate per minute at pH 7.7, 25 °C, using a system containing 0.6 mM ATP and 0.1 mM D-luciferin. The former nanomolar unit is equivalent to approximately 1.3 x 106 light units. | |
| Other Notes | Arsenate free. |
| Packaging | Sold on the basis of protein content |
| Preparation Note | Chromatographically prepared and crystallized. |
| Unit Definition | One light unit produces a biometer peak height equivalent to 0.02 μCi of 14C in PPO/POPOP cocktail. Light units measured in 50 μl assay mixture containing 5 pmol ATP and 7.5 nmol luciferin in Tris-glycine buffer, pH 7.6, at 25 °C. |
| Physical form | Lyophilized powder approximately 20% protein; balance is primarily NaCl, HEPES buffer salts, and carbohydrate. |
| Application | The reaction of this enzyme with luciferin, ATP, and O2 results in the emission of light. Luciferase can be used to detect trace amounts of ATP. Firefly luciferase is also one of the most commonly utilized reporter genes for the study of gene expression. The bioluminescent reaction catalyzed by luciferase is one of the most sensitive analytical tools for measuring gene expression. Less than or equal to one femtomole of ATP can be detected using 0.2 μg of luciferase. |
Properties
| grade | BioUltra |
| assay | ≥98% (SDS-PAGE) |
| form | lyophilized powder |
| mol wt | mol wt 120 kDa (two non-identical subunits, each containing four free thiol groups, one of which is necessary for activity) |
| composition | Protein, 10-35% E |
| measuring range | ≤1 femtomole ATP sensitivity, (using 0.2 μg of luciferase and suitably sensitive liquid scintillation counters or luminometers) |
| EQP level | Premium |
| foreign activity | ATPase ≤5 nanomolar units/mg protein |
| Nucleoside diphosphokinase ≤20 nanomolar units/mg protein | |
| storage temp. | −20°C |
Safety
Related Products
| Substrate | L6882, |
References
| reference | Stanley, P.E., A review of bioluminescent ATP techniques in rapid microbiology. J. Biolumin. Chemilumin. 4, 375, (1989) |
| Kricka, L.J., Clinical and biochemical applications of luciferases and luciferins. Anal. Biochem. 175, 14-21, (1988) | |
| Rajgopal, S., and Vijayalakshmi, M.A. Enzyme Microb. Technol. 6, 482-489, (1984) | |
| Chappelle, E.W., et al., Determination of bacterial content in fluids,. Meth. Enzymol. 57, 65-72, (1978) |
