Promega Imagine a world without 35S methionine. It's easy with Promega's TranscendTM Non-Radioactive Translation Detection Systems. Utilizing Promega's novel biotinylated lysine tRNA, these systems can be used to detect in vitro translation products after blotting with a variety of detectors, including Promega's own TranscenTM chemiluminescent substrates. No more wipe tests, no more radioactive waste disposal headaches. And results can be obtained in half the time.
One microliter of TNR translation reaction was spearated on a SDS-PAGE gel, blotted onto a PVDF membrane. The biotinylated proteins were detected with Streptavid-HRP and TranscendTM Chemiluminescent Substrates. (Lane 1 beta-galactosidase, Lanes 2 CAT and Lane 3 luciferase)
Promega has looked at the effect of biotin on enzyme and expression levels of three proteins in a reticulocyte lysate system. Two of the three (luciferase and CAT) showed no or minimal effects of biotin, while beta-galactosidase activity and expression were reduced (53% of enzymatic activity in the worst case and 70% of synthesis relative to a control reaction). Estimates of incorporation levels indicated that 25-33% of lysine residues are biotinylated in translation products that contain 4-6 lysine residues.
The kit comes in several forms: the TranscendTM Colorimetric Translation Detection System, with sufficient reagents to label 30 translation reactions and to perform colorimetric detection of biotinylated proteins on six blots; the TranscendTM Chemiluminescent Translation Detection System, with sufficient reagents to label 30 translation reactions and perform chemiluminescent detection of biotinylated proteins on six blots; or the TranscendTM tRNA alone.
For more information, contact Promega at 800-356-9526 or visit their web site at www.promega.com.