دورية أكاديمية
Biochemical Separation of Cytoplasmic and Nuclear Fraction for Downstream Molecular Analysis.
| العنوان: | Biochemical Separation of Cytoplasmic and Nuclear Fraction for Downstream Molecular Analysis. |
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| المؤلفون: | Huynh HT; Department of Medical Biochemistry and Cell biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.; Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden., Shcherbinina E; Department of Medical Biochemistry and Cell biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.; Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden., Huang HC; Department of Medical Biochemistry and Cell biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.; Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden., Rezaei R; Department of Medical Biochemistry and Cell biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.; Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden., Sarshad AA; Department of Medical Biochemistry and Cell biology, Institute of Biomedicine, University of Gothenburg, Gothenburg, Sweden.; Wallenberg Centre for Molecular and Translational Medicine, University of Gothenburg, Gothenburg, Sweden. |
| المصدر: | Current protocols [Curr Protoc] 2024 May; Vol. 4 (5), pp. e1042. |
| نوع المنشور: | Journal Article |
| اللغة: | English |
| بيانات الدورية: | Publisher: John Wiley & Sons Country of Publication: United States NLM ID: 101773894 Publication Model: Print Cited Medium: Internet ISSN: 2691-1299 (Electronic) Linking ISSN: 26911299 NLM ISO Abbreviation: Curr Protoc Subsets: MEDLINE |
| أسماء مطبوعة: | Original Publication: Hoboken, NJ : John Wiley & Sons, [2021]- |
| مواضيع طبية MeSH: | Cytoplasm*/metabolism , Cytoplasm*/chemistry , Cell Nucleus*/metabolism , Cell Nucleus*/chemistry , Cell Fractionation*/methods, Humans ; Electrophoresis, Polyacrylamide Gel ; Blotting, Western |
| مستخلص: | Biochemical fractionation is a technique used to isolate and separate distinct cellular compartments, critical for dissecting cellular mechanisms and molecular pathways. Herein we outline a biochemical fraction methodology for isolation of ultra-pure nuclei and cytoplasm. This protocol utilizes hypotonic lysis buffer to suspend cells, coupled with a calibrated centrifugation strategy, for enhanced separation of cytoplasm from the nuclear fraction. Subsequent purification steps ensure the integrity of the isolated nuclear fraction. Overall, this method facilitates accurate protein localization, essential for functional studies, demonstrating its efficacy in separating cellular compartments. © 2024 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol: Biochemical fractionation Support Protocol 1: Protein quantification using Bradford assay Support Protocol 2: SDS/PAGE and Western blotting. (© 2024 The Authors. Current Protocols published by Wiley Periodicals LLC.) |
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| فهرسة مساهمة: | Keywords: biochemical fractionation; cellular compartmentalization; cytoplasmic extraction; nuclear isolation; protein localization |
| تواريخ الأحداث: | Date Created: 20240520 Date Completed: 20240520 Latest Revision: 20240520 |
| رمز التحديث: | 20240520 |
| DOI: | 10.1002/cpz1.1042 |
| PMID: | 38767195 |
| قاعدة البيانات: | MEDLINE |
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