Homogenization of mammalian tissue

Cold Spring Harbor Protocols
Richard J Simpson

Abstract

To purify or characterize an intracellular protein, it is important to choose an efficient method for disrupting the cell or tissue that rapidly releases the protein from its intracellular compartment into a buffer that is not harmful to the biological activity of the protein of interest. One of the most widely used methods for disrupting soft tissues is homogenization. This protocol describes three processes for homogenization of animal tissues using mechanical shear: a Potter-Elvehjem glass-Teflon homogenizer, a Dounce hand homogenizer, or a handheld Waring Blendor. These methods are rapid and pose little risk to proteins other than the release of proteases from other cellular compartments. Proteolytic degradation can be minimized by the inclusion of protease inhibitors in the homogenization buffers. An optional procedure to remove mucin from tissue homogenates is also presented.

References

Jan 1, 1990·Methods in Enzymology·P Gegenheimer
Jan 1, 1990·Methods in Enzymology·J D Dignam
Jan 1, 1964·Advances in Clinical Chemistry·G B GLASS

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Citations

Apr 17, 2014·BioMed Research International·Sean C Taylor, Anton Posch
Apr 29, 2015·Molecular and Cellular Biochemistry·Olga Yu SudarkinaLyudmila V Dergunova
Aug 4, 2010·Cold Spring Harbor Protocols·Hong Ji
Jul 3, 2019·Cold Spring Harbor Protocols·James DeCaprio, Thomas O Kohl
Oct 5, 2017·Expert Review of Proteomics·Manish MishraAldrin V Gomes
Mar 21, 2018·Biochemistry·Tomohiro KimuraRichard M Epand
Nov 3, 2010·Cold Spring Harbor Protocols·Richard J Simpson

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