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Preparation of representative homogenates of biological tissues: effect of salt on protein extraction
G C Upreti1, C Davis, J Oliver
1Animal Research Group, Ruakura Agricultural Centre, Hamilton, New Zealand.
A technique for the preparation of representative homogenates of tissues is described. For most soft tissues and biological fluids (liver, kidney, semen) more than 80% of protein was recovered after homogenization in isotonic buffer. With tissues that are harder to homogenize, such as heart, spleen, and skeletal muscle, however, only approximately 40-50% of protein was extracted in this medium. Inclusion of salt or salt plus detergent during the homogenization increased recovery of the protein to levels close to those recorded with soft tissues. The increase represented a true recovery and was not an artifact induced by salt/detergent. This situation is parallel to previously reported results for lipid-rich biological tissues.
A technique for the preparation of representative homogenates of tissues is described. For most soft tissues and biological fluids (liver, kidney, semen) more than 80% of protein was recovered after homogenization in isotonic buffer. With tissues that are harder to homogenize, such as heart, spleen, and skeletal muscle, however, only approximately 40-50% of protein was extracted in this medium. Inclusion of salt or salt plus detergent during the homogenization increased recovery of the protein to levels close to those recorded with soft tissues. The increase represented a true recovery and was not an artifact induced by salt/detergent. This situation is parallel to previously reported results for lipid-rich biological tissues.