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| 1. |
Salerno, M.S., Thomas, M., Forbes, D., Watson, T., Kambadur, R., and Sharma, M.
(2004)
Molecular analysis of fiber type-specific expression of murine myostatin promoter.
Am. J. Physiol., Cell Physiol.
287
,
1031-1040
.
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Notes:
In this article, a 2.5- and a 1.7-kb fragment of the myostatin promoter were amplified by PCR and cloned into the pGEM®-T Easy vector. These clones were then subcloned into the pGL3-Basic Vector. Deletion mutants of the constructs were made and used to transient transfect mouse muscle C2C12 cells. The Luciferase Assay System was then used to analyze transfectants. The researchers also injected luciferase-reporter constructs into mouse muscle. Luciferase activity in the mouse muscle was examined by grinding isolated muscles in liquid nitrogen and resuspending them in Cell Culture Lysis Buffer. Ten micron cryosections of the muscles were also used in immunohistochmical staining experiments. For these experiments, the researchers used a 1:50 dilution of Promega’s polyclonal anti-luciferase antibody, a secondary antibody and tertiary fluorescein-labeled conjugate. The slides were counter stained with DAPI.
(0003147) |
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Products: Anti-Luciferase pAb | Luciferase 1000 Assay System | Luciferase Assay System | Luciferase Assay System Freezer Pack | Luciferase Assay System with Reporter Lysis Buffer | Luciferase Assay System, 10-Pack | Luciferase Cell Culture Lysis 5X Reagent | pGEM®-T Easy Vector System I | pGEM®-T Easy Vector System II | pGL3-Basic Vector |
| 2. |
Ashley, W.W. Jr, and Russell, B.
(2000)
Tenotomy decreases reporter protein synthesis via the 3'-untranslated region of the beta-myosin heavy chain mRNA.
Am. J. Physiol., Cell Physiol.
279
,
C257-65
.
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Notes:
In a rat model of muscle atrophy, the myosin heavy chain 3'-untranslated region mediates a decrease in myosin heavy chain expression. To quantitate this effect, luciferase was placed under the control of either the myosin heavy chain or SV40 3'-UTR in the pGL3 Control Vector to show that luciferase expression decreased 63% in this model compared to controls. Luciferase assay were performed using one of Promega's luciferase substrates (not specified). For luciferase assays, soleus muscle was ground to a fine powder in liquid nitrogen in a cell lysis buffer (0.5% Nonidet® P-40 (NP-40), 10 mM HEPES, 3 mM MgCl2, 40 mM KCl, 2 mM dithiothreitol (DTT), 0.5 mM PMSF, and a protease inhibitor cocktail. The tissue was homogenized on ice, centrifuged at 3,000 rpm at 4°C to remove debris, and stored at 80°C. For immunohistochemistry, tissue was frozen, serial sectioned into 10µm slices, fixed with 4% paraformaldehyde, and stained with the Anti-Luciferase pAb.
(0002449) |
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Products: Anti-Luciferase pAb |