Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase
A unique combined luminescence assay for firefly (Photinus pyralis) luciferase and β-galactosidase (β-gal) reporter gene products is described. Luciferase and β-gal activities are determined with the same aliquot of cell lysate prepared from cells contransfected with both reporter genes, thereby red...
Saved in:
| Main Authors: | , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Taylor & Francis Group
1996-09-01
|
| Series: | BioTechniques |
| Online Access: | https://www.future-science.com/doi/10.2144/96213pf01 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1850152337061969920 |
|---|---|
| author | Chris S. Martin Patricia A. Wight Anna Dobretsova Irena Bronstein |
| author_facet | Chris S. Martin Patricia A. Wight Anna Dobretsova Irena Bronstein |
| author_sort | Chris S. Martin |
| collection | DOAJ |
| description | A unique combined luminescence assay for firefly (Photinus pyralis) luciferase and β-galactosidase (β-gal) reporter gene products is described. Luciferase and β-gal activities are determined with the same aliquot of cell lysate prepared from cells contransfected with both reporter genes, thereby reducing manual labor and increasing experimental accuracy. With the Dual-LightTM assay system, luciferase activity is measured first with an enhanced luciferase assay, followed by quantitation of β-gal with Galacton-PlusTM chemiluminescent substrate and Sapphire-IITM enhancer. Highly sensitive detection of luciferase (2 fg) and β-gal (8 fg) is achieved with a dynamic range over seven orders of magnitude of enzyme concentration. Comparative analysis of both independent and combined (Dual-Light) detection methods for cells contransfected with luciferase and β-gal reporter genes is also described. |
| format | Article |
| id | doaj-art-bfc5ce0297d9473fbdc4fb4ce0d55938 |
| institution | OA Journals |
| issn | 0736-6205 1940-9818 |
| language | English |
| publishDate | 1996-09-01 |
| publisher | Taylor & Francis Group |
| record_format | Article |
| series | BioTechniques |
| spelling | doaj-art-bfc5ce0297d9473fbdc4fb4ce0d559382025-08-20T02:26:00ZengTaylor & Francis GroupBioTechniques0736-62051940-98181996-09-0121352052410.2144/96213pf01Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-GalactosidaseChris S. Martin0Patricia A. Wight1Anna Dobretsova2Irena Bronstein31Tropix, Inc., Bedford, MA1Tropix, Inc., Bedford, MA1Tropix, Inc., Bedford, MA1Tropix, Inc., Bedford, MAA unique combined luminescence assay for firefly (Photinus pyralis) luciferase and β-galactosidase (β-gal) reporter gene products is described. Luciferase and β-gal activities are determined with the same aliquot of cell lysate prepared from cells contransfected with both reporter genes, thereby reducing manual labor and increasing experimental accuracy. With the Dual-LightTM assay system, luciferase activity is measured first with an enhanced luciferase assay, followed by quantitation of β-gal with Galacton-PlusTM chemiluminescent substrate and Sapphire-IITM enhancer. Highly sensitive detection of luciferase (2 fg) and β-gal (8 fg) is achieved with a dynamic range over seven orders of magnitude of enzyme concentration. Comparative analysis of both independent and combined (Dual-Light) detection methods for cells contransfected with luciferase and β-gal reporter genes is also described.https://www.future-science.com/doi/10.2144/96213pf01 |
| spellingShingle | Chris S. Martin Patricia A. Wight Anna Dobretsova Irena Bronstein Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase BioTechniques |
| title | Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase |
| title_full | Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase |
| title_fullStr | Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase |
| title_full_unstemmed | Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase |
| title_short | Dual Luminescence-Based Reporter Gene Assay for Luciferase andβ-Galactosidase |
| title_sort | dual luminescence based reporter gene assay for luciferase andβ galactosidase |
| url | https://www.future-science.com/doi/10.2144/96213pf01 |
| work_keys_str_mv | AT chrissmartin dualluminescencebasedreportergeneassayforluciferaseandbgalactosidase AT patriciaawight dualluminescencebasedreportergeneassayforluciferaseandbgalactosidase AT annadobretsova dualluminescencebasedreportergeneassayforluciferaseandbgalactosidase AT irenabronstein dualluminescencebasedreportergeneassayforluciferaseandbgalactosidase |