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Increase Protein Activity and Yield

ActivePro In Vitro Translation Kit

Higher yields of protein than reticulocyte lysate systems
Fully compatible with eukaryotic protein translation
Higher biological activity than other E. coli lysate systems
Easy to use

For researchers studying protein function and protein-protein interactions, Ambion's ActivePro In Vitro Translation Kit is the ideal tool. ActivePro uses an E. coli lysate to synthesize high yields of functional protein for studies where properly folded active protein is a necessity. Furthermore, research data show that ActivePro is capable of efficiently synthesizing multiple proteins in a single reaction--an important feature for protein interaction studies.

Higher Yields than Retic Systems
Translation of a sequence in ActivePro typically yields greater than 1000-fold more protein than coupled reticulocyte lysate systems. What's more, ActivePro synthesizes protein at a very high rate (several hundred micrograms per hour per ml of reaction), with the reaction 80% complete in just 1 hour. In contrast, typical coupled retic kits require 1.5 hours longer incubation. An efficient energy regeneration system and a large supply of amino acids included in the ActivePro Kit support faster protein synthesis and higher yields.

Coexpression of Multiple Proteins
Comparison data with competing kits have already illustrated ActivePro's superior ab ility to synthesize protein with higher biological activity from both prokaryotic and eukaryotic transcripts. Now, more recent data shows ActivePro's ability to synthesize higher yields of multiple proteins in the same reaction. Two viral proteins, protein A and protein B, were synthesized in separate reactions using ActivePro and two other commercial E. coli lysate systems. Figure 1 illustrates the 2-3 fold greater yield of both proteins with ActivePro compared with the competition.

Figure 1. Expression Yields from Two Viral Proteins. DNA templates for protein A and protein B were transcribed/translated in vitro using Ambion's ActivePro Kit (1 hour reaction at 37C) and 2 other commercially available E. coli lysate kits, according to the recommended protocol. Each reaction included 20 Ci 35S-labeled methionine. Yield was assayed by TCA precipitation, and, post-acetone precipitation, 5 l of the reaction was analyzed by SDS-PAGE.

This same translation efficiency also holds true when coexpressing multiple proteins in the same reaction (Figure 2). Cell-free coexpression of two plasmids in the same reaction is useful in interaction studies such as ligand binding experiments, receptor studies, and protein-protein interaction studies. ActivePro has the proven ability not only to coexpress multiple proteins, but to do so with higher yields than the competition.

Figure 2. Cell-free Coexpression of Multiple Proteins. The DNA templates for protein A and protein B (cloned into the included vector) were coexpressed in the same reaction using the ActivePro Kit (1 hour reaction at 37C). The final reaction mixture was analyzed by SDS-PAGE.

Easy to Use
ActivePro is simple to use. You provide only a labeled amino acid (optional) and a DNA template that contains your gene of interest, a T7 promoter, ribosome binding site, and T7 terminator. The DNA template can be easily constructed by inserting your gene of interest into the multiple cloning site of Ambion's control plasmid included in the kit. In a single tube, simply premix the reagents included in the kit and add your DNA template, labeled amino acid, if desired, and incubate at 37C for 1 hour. The resulting biologically active protein is then ready for further functional analysis.

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


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