What is TK Renilla vector?
The pRL Vectors are wild-type Renilla luciferase (Rluc) control reporter vectors. The HSV-thymidine kinase promoter (pRL-TK) is relatively weak and may be particularly useful in providing neutral constitutive expression of the Renilla luciferase control reporter.
What is pRL TK?
The thymidine kinase promoter-Renilla luciferase reporter plasmid (pRL-TK) is commonly used as a control for transfection efficiency in the Dual-Luciferase Reporter Assay System.
What is Renilla luciferase used for?
The Renilla luciferase gene is commonly used as an internal control in luciferase-based reporter gene assays to normalize the values of the experimental reporter gene for variations that could be caused by transfection efficiency and sample handling.
What is dual luciferase reporter assay?
The Dual-Luciferase® Reporter (DLR™) Assay System provides an efficient means of performing two reporter assays. In the DLR™ Assay, the activities of firefly (Photinus pyralis) and Renilla (Renilla reniformis or sea pansy) luciferases are measured sequentially from a single sample.
How can you detect luciferase expressing cells?
To measure luciferase activity of the cell lysates, you will need a multiwell plate or a tube containing cell lysates and a luminometer. This apparatus detects the photon emission produced from the luciferase reaction and the unit of measurement comes out as relative light units (RLU).
What is firefly luciferase assay?
The Luciferase Assay System is an extremely sensitive and rapid reagent for quantitation of firefly luciferase. Linear results are seen over at least eight orders of magnitude of enzyme concentration, and less than 10–20 moles of luciferase can be measured under optimal conditions.
Can luciferase be scanned?
The bioluminescence emitted can be detected and amplified using specialized cameras with highly sensitive detection systems , revealing the sites and levels of luciferase expression and activity from within a living animal.
Why is luciferase the best reporter?
Luciferase-based assays are better than other reports because of the following advantages: Quick and real-time measurement. Exceptionally high sensitivity than fluorescent reporters like GFP (10- to 1,000 fold) Range of measurement is wide and dynamic.
What is the difference between firefly and Renilla luciferase?
What is the Difference Between Firefly and Renilla Luciferase? Firefly luciferase assays uses luciferin in the presence of oxygen, ATP and magnesium to produce light (Green/Yellow, 550-70 nM), while Renilla luciferase assays (SCT153) requires only coelenterazine and oxygen to produce light (Blue, 480 nM).
Is luciferin toxic to humans?
Luciferin Toxicity The small size of luciferin also makes it a poor antigen and immune responses to luciferin are unlikely. Luciferin is able to pass the blood brain barrier, the blood placenta barrier and the blood testis barrier, toxicity appears low.
Is firefly luciferase secreted?
However, Cypridina and Gaussia luciferases are secreted; therefore, Pierce Cypridina-Firefly and Gaussia-Firefly dual-spectral luciferase kits are compatible with time-course assays.
Why is it called luciferin?
“Luciferin” got its name from the Latin word “lucifer” (meaning “light-bearing”), which is also a source of the word that is sometimes used as a name of the devil.
What are PRL vectors?
The pRL Vectors are wild-type Renilla luciferase ( Rluc) control reporter vectors. The pRL Vectors, which provide constitutive expression of Renilla luciferase, can be used in combination with a firefly luciferase vector to cotransfect mammalian cells.
Is the early SV40 enhancer/promoter region suitable for co-reporter applications?
The early SV40 enhancer/promoter region (pRL-SV40) and the CMV immediate early enhancer/promoter region (pRL-CMV) typically provide high-level transcription and, therefore, may be less suitable for co-reporter applications involving experimental vectors with robust regulatory elements.
How can I use the PRL-null vector as a control?
The pRL-null Vector lacks a promoter and offers a multiple cloning site to insert a regulatory region of interest. Alternatively, use the vector as a control without a promoter element if needed for your experimental assay.
Is the HSV-thymidine kinase promoter (PRL-TK) useful for Renilla luciferase control?
The HSV-thymidine kinase promoter (pRL-TK) is relatively weak and may be particularly useful in providing neutral constitutive expression of the Renilla luciferase control reporter.