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Translate High Yields of Active Protein

ActivePro In Vitro Translation Kit


Translation products have higher biological activity than those from other E. coli lysate systems

Efficient synthesis of a wide range of prokaryotic and eukaryotic proteins

Fast translation and high protein yield

Easy to use

You really can have it all with the the ActivePro In Vitro Translation Kit--high yields, active protein, and short reaction time. Get more full-length, active protein from both prokaryotic and eukaryotic templates.


High Yields of Active Protein
There are several in vitro translation kits available commercially, but only one has it all -- high yields of biologically active protein in the shortest amount of time. The ActivePro In Vitro Translation Kit is a coupled transcription/translation system capable of expressing prokaryotic and eukaryotic sequences in 1-2 hours. It is a cell-free, scalable, simple to use kit, that is ideal for functional assays, protein interaction studies, and expression screenings. Compared to other E. coli lysate systems, ActivePro generates proteins with higher biological activity, in comparable yields.

Figure 1 compares the yield and activity of proteins expressed using the ActivePro Kit with those from two other E. coli lysate kits. The experiment includes both a prokaryotic (CAT) and a eukaryotic (GFP) protein. The data demonstrate ActivePro's superior ability to translate high yields of f 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 dase by colorimetric ONGP hydrolysis assay


Human Protein Expression
ActivePro's versatility with a wide range of proteins is further demonstrated by the expression of human templates--Caspase3, MAPK9, and Casein Kinase. The results from expressing these human proteins using the ActivePro Kit are shown in Figure 3.

Figure 3. Complex Human Proteins are Translated With the Same Yield and Efficiency as other Types . (A) DNA templates (1-2 g) for three human proteins were in vitro transcribed/translated using 20 Ci 35S-labeled methionine for 1 hour at 37C according to the ActivePro protocol. 5 l of each reaction were assessed by SDS-PAGE gel and phosphorimaging. (B) Table summarizing data on protein expression yields (calculated as described in Figure 2).


Make the Switch Today
Whether you are currently using an E. coli or a reticulocyte lysate translation system, making the switch to Ambion's ActivePro In Vitro Translation Kit can give you higher yields of more active protein in the shortest amount of time.


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Ordering Information
Cat# Product Name Size 1295 ActivePro In Vitro Translation Kit 25 rxns
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