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Chapters
- 00:05Title
- 00:35Site-directed Mutagenesis
- 01:45Transformation and Screening for the Desired Clone
- 02:35pcDNA3.1/GLA and pcDNA3.1/GAA Plasmid Transfection and Treatment of HEK293H
- 04:00Cell Harvest
- 04:43Homogenization by Freezing and Thawing and BCA Assay
- 06:17Enzyme Activity Measurement with Artificial 4-Methylumbeliferyl Substrate
- 08:19Results: Enzyme Activity Measurement Mutagenized 3 α-galactosidase and 3 Acid α-glucosidase
- 09:35Conclusion
Há uma demanda para fazer pré-clínicos de teste para uma novela classe de drogas "órfão" chamado acompanhantes farmacológicos podem ser reproduzidos, rápido e eficiente. Desenvolvemos um ensaio de baseado em cultura celular altamente padronizado, simples e versátil para tela para pacientes elegíveis, bem como drogas de romance acompanhante farmacológica.
Tags
In VitroEnzyme MeasurementPharmacological ChaperoneFabry DiseasePompe DiseasePhenotypic AssessmentAllelic VarianceLysosomal Storage DiseasesPrognostic OutcomesTherapeutic DecisionsSite-directed MutagenesisGLA GeneGAA GenePrimer DesignPCRDpn1E.coliPlasmid PurificationSpectrophotometerHEK293H CellsCell Transfection
