Luciferase Reporter Vector back

 

Wide array of carefully optimized reagents designed to induce transcription factors.

 

With Panomics Luciferase Reporter Vectors, you can monitor and even quantify the binding activity of transcription factors. Each Reporter Vector contains a cis-acting DNA binding element that is recognized by a specific TF. Binding at this site results in the expression of firefly luciferase, an enzyme capable of catalyzing a powerful bioluminescent reaction. Light emitted from the chemical reaction is directly proportional to the amount of expressed enzyme and thus the binding activity of the targeted TF.

 

Like our EMSA gel-shift kits, the Panomics Reporter Vectiors are ideal for validating the results obtained using Protein/DNA Arrays. But while EMSA only allows in vitro verification of those results, the use of the Luciferase Reporter Vectors allows TF activity to be monitored in living cells. Don't settle for answers from the test tube; investigate TF binding where it happens!

 

51 vectors now available!

 

·         Quantitative. Use luciferase activity to measure TF binding

·         Flexible. Use for in vivo assays

·         Rapid. Get results in hours

·         Stable. Store for up to one year

·         Safe. No radioactivity required

 

 

Reporter Construct

 

The Luciferase Reporter Vectors have been specially constructed to report the binding activity of a single TF. Multiple copies of the cis-acting enhancer element have been inserted into each vector upstream of a minimal TA promoter and the TATA box from the Herpes simplex virus thymidine kinase promoter. This promoter sequence drives expression of the luciferase gene (luc). The backbone of the vector contains an ampicillin resistance gene for cloning purposes, an origin of replication, and an f1 origin for single-stranded DNA production.

 

 

 

 

Using the Luciferase Assay to Monitor TF Activity

 

As a means of measuring promoter response in cells, the luciferase assay is simple, straightforward, and very effective. The reporter vector is first transfected into cells. After a limited amount of time, the cells are lysed and the substrate of luciferase, luciferin, is introduced into the cellular extract along with Mg and excess ATP.

Under these conditions, luciferase enzyme expressed by the reporter vector will catalyze the oxidative carboxylation of luciferin. The luminescence from this chemical reaction can be read and quantified by a luminometer or scintillation counter. The amount of light detected from the cell lysate correlates directly with the binding activity of the transcription factor.

 

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