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Protein Extraction From Mammalian Tissues

  • Choogon Lee
Part of the Methods in Molecular Biology™ book series (MIMB, volume 362)

Abstract

For Western blotting and coimmunoprecipitation (coIP), protein samples must be extracted from tissues. The protocol described in this chapter has been used to extract clock proteins from mammalian tissues as diverse as liver, kidney, and brain. The extraction protocol is mild enough to be used for coIP as well as Western blotting. Simply, clock proteins are extracted from tissues by freezing and thawing, and homogenizing with a handheld homogenizer. This procedure extracts most (>90%) of the clock proteins from mammalian tissues.

Key Words

Clock proteins Western blotting coimmunoprecipitation extraction mammalian tissue 

References

  1. 1.
    Lee, C., Etchegaray, J. P., Cagampang, F. R. A., Loudon, A. S. I., and Reppert, R. M. (2001) Posttranslational mechanisms regulate the mammalian circadian clock. Cell 107, 855–867.CrossRefPubMedGoogle Scholar
  2. 2.
    Braford, M. M. (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal. Biochem. 72, 248–254.CrossRefGoogle Scholar

Copyright information

© Humana Press Inc. 2007

Authors and Affiliations

  • Choogon Lee
    • 1
  1. 1.Department of Biomedical SciencesFlorida State University College of MedicineTallahassee

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