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Anal. Chem. 67, 4260-4294. Expression immunoassay. Antigen quantitation using antibodies labeled with enzyme-coding DNA fragments. 1995

Christopoulos, T.K. and Chiu, N.H.L.

Notes: These researchers use a nucleic acid fragment/complete expression unit as a label in immunoassays. For a model, firefly luciferase DNA was used. The immunoreaction is carried out with biotinylated antibodies, which are attached to the DNA via the streptavidin molecule. The DNA is then transcribed and translated using the TNT® T7 Wheat Germ Extract System. The reaction of luciferase produces measurable luminescence that is proportional to the number of antigen molecules. (1903)

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Biotechniques 18, 244-248. Rapid screening of open reading frames by protein synthesis with an in vitro transcription and translation assay. 1995

Switzer, W.M., Heneine, W.

Notes: This article details a method for using the TNT® Coupled Reticulocyte Lysate System to rapidly screen ORFs (using 1/2 the reaction components per assay) directly from bacterial colonies without performing plasmid preps. (0286)

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Eur. J. Biochem. 228, 222-228. Retinoid-X receptor-alpha-independent binding of vitamin D receptor to its response element from human osteocalcin gene. 1995

Jääskeläinen, T., Itkonen, A., and Mäenpää, P.H.

Notes: Retinoid X-receptor was transcribed and translated in vitro using the TNT® Coupled Wheat Germ Extract System. The protein was used for EMSAs with Ap-1 and VDR responsive regions from the human osteocalcin gene. Both half sites of the VDRE are required for VDR-RXRalpha binding, suggesting that RXRalpha is not the physiological accessory factor for VDR-VDRE interactions. The RXRalpha protein was also produced in the presence of [35S]methionine and analyzed by SDS-PAGE. (1805)

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Nucl. Acids Res. 23, 588-594. The DNA binding domains of the yeast Gal4 and human c-Jun transcription factors interact through the zinc-finger and bZIP motifs. 1995

Sollerbrant, K., Akusjarvi, G., Linder, S. and Svensson, C.

Notes: c-Jun, c-Jun-bZIP and cJun-TAD were synthesized using the TNT® SP6 Coupled Wheat Germ Extract System. The proteins were used in GST binding assays with GSTGal4, GSTGal4(1-147) and GSTGal4/CR1. These assays show that Gal4 and c-Jun can physically interact in vitro. Gal4(1-147) increases the binding of c-Jun to a consensus AP1 DNA binding site in EMSAs. (1813)

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Plant Mol. Biol. 29, 317-330. The in vitro assembly of the NADPH-protochlorophyllide oxidoreductase in pea chloroplasts. 1995

Dahlin, C. , Sundqvist, C. , Timko, M. P.

Notes: [3H]leucine-labeled pchlide reductase was produced in the TNT® Coupled Wheat Germ Extract System. These proteins were used in membrane assembly assays in chloroplast lysates. The results indicate that the pchlide reductase is a peripheral protein located on the stromal side of the membrane. (1294)

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Biochem. Soc. Trans. 22, 413S. Nuclear expression of mitochondrial genes implicated in human encephalomyopathies. 1994

Sutherland, L., Davidson, J., Jacobs, H. T.

Notes: This paper discusses the use of the TNT® Coupled Reticulocyte Lysate System to express mitochondrial genes in the mammalian cytosol and check for protein expression. (0324)

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J. Virol. 67, 7612-7617. NF-kappaB p100 is one of the high-molecular weight proteins complexed with the v-Rel oncoprotein in transformed chicken spleen cells 1993

Sif, S. and Gilmore, T.D.

Notes: v-Rel, c-Rel, p105 and p100 were expressed individually and together using the TNT® SP6 Coupled Wheat Germ Extract System. The cotranslation products of all four proteins were immunoprecipitated with anti-v-Rel antiserum. All four proteins were found in the immunoprecipitates, although p105 and p100 are not recognized by v-Rel antiserum. Thus, p105 and p100 are immunoprecipitated by their association with the complex. (1833)

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