Abstract
Cell fractionation is a method used to study different cellular events like protein translocation and sequestration by disrupting cells and fractionating their contents, thus allowing an enrichment of the protein of interest. Using different concentrations of sucrose or detergent buffer formulations in combination with centrifugations, the cell fractions are separated based on their density and size.
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References
Claude A (1946) Fractionation of mammalian liver cells by differential centrifugation: I. Problems, methods, and preparation of extract. J Exp Med 84:51–59
Claude A (1946) Fractionation of mammalian liver cells by differential centrifugation: II. Experimental procedures and results. J Exp Med 84:61–89
de Duve C, Pressman BC, Gianetto R et al (1955) Tissue fractionation studies. 6. Intracellular distribution patterns of enzymes in rat-liver tissue. Biochem J 60:604–617
Blobel G, Potter VR (1966) Nuclei from rat liver: isolation method that combines purity with high yield. Science 154:1662–1665
Bronfman M, Loyola G, Koenig CS (1998) Isolation of intact organelles by differential centrifugation of digitonin-treated hepatocytes using a table Eppendorf centrifuge. Anal Biochem 255:252–256
Srinivas KS, Chandrasekar G, Srivastava R et al (2004) A novel protocol for the subcellular fractionation of C3A hepatoma cells using sucrose density gradient centrifugation. J Biochem Biophys Methods 60(1):23–27
Suzuki K, Bose P, Leong-Quong RY et al (2010) REAP: a two minute cell fractionation method. BMC Res Notes 3:294
Graham JM, Rickwood (1997) Subcellular fractionation a practical approach. IRL Press Oxford University Press, Oxford
Capece D, Verzella D, Di Francesco B et al (2020) NF-κB and mitochondria cross paths in cancer: mitochondrial metabolism and beyond. Semin Cell Dev Biol 98:118–128
Begalli F, Bennett J, Capece D et al (2017) Unlocking the NF-κB conundrum: embracing complexity to achieve specificity. Biomedicine 5(3):50
Bennett J, Capece D, Begalli F et al (2018) NF-κB in the crosshairs: rethinking an old riddle. Int J Biochem Cell Biol 95:108–112
Baud V, Karin M (2009) Is NF-κB a good target for cancer therapy? Hopes and pitfalls. Nat Rev Drug Discov 8(1):33–40
Acknowledgments
The work was supported by the MIUR PRIN grant n° 2017WLKYAM_1 to F.Z.
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Vecchiotti, D. et al. (2021). Biochemical Methods to Analyze the Subcellular Localization of NF-κB Proteins Using Cell Fractionation. In: Franzoso, G., Zazzeroni, F. (eds) NF-κB Transcription Factors. Methods in Molecular Biology, vol 2366. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1669-7_2
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DOI: https://doi.org/10.1007/978-1-0716-1669-7_2
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-1669-7
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