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Translate High Yields of Biologically Active Proteins

ion With ActivePro Surpasses Competitors
To compare the efficiency of ActivePro with two competing systems, five human templates were expressed using the manufacturers protocols. Both yields (Figure 3) and biological activities of the reaction products (Figure 4) were assayed. ActivePro gave the highest yields for all five of the proteins translated. And in all but one case (DHFR), ActivePro-translated proteins showed the highest enzymatic activity.

Figure 3. Comparison of Protein Yields Across E. coli Lysate Kits. DNA templates for 5 human proteins were in vitro transcribed/translated using Ambions ActivePro Kit and two other commercially available kits, according to protocol. The proteins analyzed were dihydrofolate reductase (DHFR), adenylate kinase 1 (AK1), carbonic anhydrase 2 (CA2), caspase 3 (CASP3), and casein kinase 1 (CK1). (A) Quantitated protein yields as determined by TCA precipitation. (B) Autoradiographs of the 35S-labeled proteins.

Figure 4. Comparison of Biological Activity Across E. coli Lysate Kits. The proteins as produced from Figure 3 were tested in functional assays. Relative activities from protein preparations synthesized using two commercially available kits were compared to that from the ActivePro Kit. Dihydrofolate reductase (DHFR): Activity was measured with a standard DHFR assay, which follows the rate of conversion of dihydrofolate and NADPH to tetrahydrofolate and NADP+ through the successive decrease of absorbance of NADPH at 340 nm at 30C [1]. Adenylate kinase 1 (AK1): Activity was measured by a luciferase-based ATP bioluminescen
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