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Translate High Yields of Active Protein

ull-length protein with a higher level of biological activity than competitor's kits. ActivePro's ability to transcribe/translate both eukaryotic and prokaryotic templates over a broad size range is summarized in Figure 2.

Figure 1. Protein Yield, Quality, and Activity Comparison of E. coli Lysate Kits. (A) Autoradiograph of 35S-labeled GFP and CAT proteins that were in vitro transcribed/translated using Ambion's ActivePro Kit and two other commercially available kits, according to protocol. 5 l aliquots of each reaction were assessed by SDS-PAGE and phosphorimaging. (B) Aliquots from each reaction were assessed for protein activity by either measuring GFP fluorescence on a SPECTRAmax GEMINI XS Fluorometer (Molecular Devices), or using the method described by Shaw [1] to evaluate CAT activity.

Figure 2. ActivePro Transcribes/Translates Both Prokaryotic and Eukaryotic Proteins Over a Broad Size Range. (A) DNA templates (1-2 g) for several prokaryotic and eukaryotic proteins were in vitro transcribed/translated with the ActivePro Kit in reactions that included 20 Ci 35S-labeled methionine. 5 l aliquots of each reaction were assessed by SDS-PAGE and phosphorimaging. (B) Table summarizing protein yield and activity data with the ActivePro Kit. Incorporated 35S-methionine was measured by TCA precipitation and total protein yields were calculated. CAT activity was measured using methods described by Shaw [1], Rhodanese activity by the colorimetric assay described by Sorbo [2], Luciferase activity by luminescence, GFP activity by fluorescence, and -galactosi
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