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Determination of nuclear transcription factor activity using a modified electrophoretic mobility shift assay
Wang, Yong; Huang, Wenhua; Peng, Daizhi; Hu, Yuanbin.
  • Wang, Yong; Third Military Medical University. Southwest Hospital. Institute of Burn Research; State Key Laboratory of Trauma; Burns and Combined Injury. Chongqing. CN
  • Huang, Wenhua; Third Military Medical University. Southwest Hospital. Institute of Burn Research; State Key Laboratory of Trauma; Burns and Combined Injury. Chongqing. CN
  • Peng, Daizhi; Third Military Medical University. Southwest Hospital. Institute of Burn Research; State Key Laboratory of Trauma; Burns and Combined Injury. Chongqing. CN
  • Hu, Yuanbin; Third Military Medical University. Southwest Hospital. Institute of Burn Research; State Key Laboratory of Trauma; Burns and Combined Injury. Chongqing. CN
Braz. arch. biol. technol ; 51(4): 667-672, June-Aug. 2008. ilus
Article in English | LILACS | ID: lil-622674
ABSTRACT
In this work, several major procedures of the electrophoretic mobility shift assay (EMSA) were modified including swift extraction of the nucleic protein, labeling of the probe and radioautography. The modified assay required shorter time, simplified the nucleic protein extraction, increased the radioactivity of the labeling probe, skipped the tedious process of gel drying, and produced clear images. Its results were comparable, reproducible and stable. It thus has merited for wide application.
RESUMO
A determinação da alteração na mobilidade eletroforética (EMSA), o método de mais ampla utilização para o estudo das interações proteínaácidos nucléicos, é tediosa e difícil de dominar. De acordo com os protocolos dacumentados e com base em nossa prática, nós modificamos os diversos processos principais dessa determinação incluindo no que diz respeito a extração de proteiínas nucleicas, marcação das provas e radioautografia. A determinação modificada requer menor tempo, simplifica a extração de ácidos nucleicos, eleva a radioatividade da prova marcada, evita o processo tedioso de secagem do gel e produz claras imagens. Seus resultados são comparáveis, reproduzíveis e estáveis, merecendo, desse modo, ampla aplicação.


Full text: Available Index: LILACS (Americas) Type of study: Practice guideline Language: English Journal: Braz. arch. biol. technol Journal subject: Biology Year: 2008 Type: Article / Project document Affiliation country: China Institution/Affiliation country: Third Military Medical University/CN

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Full text: Available Index: LILACS (Americas) Type of study: Practice guideline Language: English Journal: Braz. arch. biol. technol Journal subject: Biology Year: 2008 Type: Article / Project document Affiliation country: China Institution/Affiliation country: Third Military Medical University/CN