دورية أكاديمية
Cell-free expression of protein kinase a for rapid activity assays.
| العنوان: | Cell-free expression of protein kinase a for rapid activity assays. |
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| المؤلفون: | Leippe DM; Research and Development, Promega Corporation, 2800 Woods Hollow Road, Madison, WI, 53711, USA. Email: donna.leippe@promega.com., Zhao KQ, Hsiao K, Slater MR |
| المصدر: | Analytical chemistry insights [Anal Chem Insights] 2010 May 19; Vol. 5, pp. 25-36. Date of Electronic Publication: 2010 May 19. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: SAGE Publications Country of Publication: United States NLM ID: 101463741 Publication Model: Electronic Cited Medium: Internet ISSN: 1177-3901 (Electronic) Linking ISSN: 11773901 NLM ISO Abbreviation: Anal Chem Insights Subsets: PubMed not MEDLINE |
| أسماء مطبوعة: | Publication: <2016-2018> : Thousand Oaks, CA : SAGE Publications Original Publication: Auckland : Libertas Academica |
| مستخلص: | Functional protein analysis often calls for lengthy, laborious in vivo protein expression and purification, and can be complicated by the lack of stability of the purified protein. In this study, we demonstrate the feasibility of a simplified procedure for functional protein analysis on magnetic particles using cell-free protein synthesis of the catalytic subunit of human cAMP-dependent protein kinase as a HaloTag((R)) fusion protein. The cell-free protein synthesis systems provide quick access to the protein of interest, while the HaloTag technology provides efficient, covalent protein immobilization of the fusion protein, eliminating the need for further protein purification and minimizing storage-related stability issues. The immobilized cPKA fusion protein is assayed directly on magnetic beads and can be used in inhibitor analyses. The combination of rapid protein synthesis and capture technologies can greatly facilitate the process of protein expression and activity screening, and therefore, can become a valuable tool for functional proteomics studies. |
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| فهرسة مساهمة: | Keywords: HaloTag; PKA; cell-free expression; in vitro translation; kinase; magnetic particles; protein immobilization |
| تواريخ الأحداث: | Date Created: 20100604 Date Completed: 20110714 Latest Revision: 20211020 |
| رمز التحديث: | 20240628 |
| مُعرف محوري في PubMed: | PMC2879223 |
| DOI: | 10.4137/aci.s4732 |
| PMID: | 20520741 |
| قاعدة البيانات: | MEDLINE |
Full Text Finder | تدمد: | 1177-3901 |
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| DOI: | 10.4137/aci.s4732 |