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Western Blotting Analysis of CCN Proteins in Calcified Tissues

Protocol
First Online:
Part of the Methods in Molecular Biology book series (MIMB, volume 1489)

Abstract

Western blotting is widely used for protein analysis. We routinely perform such analysis for evaluating the production levels of CCN family proteins in a variety of cells under various conditions. In this chapter, we describe our Western blotting protocol to estimate protein production profiles of CCN family members after having assessed the specificity of the antibodies against each CCN member protein to ensure no cross-reaction with other CCN member proteins.

Key words

Amino acid homology Conserved module Member-specific antibody Cross-reaction Redundancy of CCN members 
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Notes

Acknowledgement

This work was supported by JSPS KAKENHI Grant Numbers JP24390415 and JP15H05014 to M.T., JP25462886 to S.K., JP26462984 to H.K., and The Science Research Promotion Fund of The Promotion and Mutual Aid Corporation for Private Schools of Japan to H.K. The authors gratefully thank Ms. Yoshiko Miyake for her secretary assistance.

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  1. 1.Department of Oral Biochemistry, Division of Oral Structure, Function, and DevelopmentAsahi University School of DentistryGifuJapan
  2. 2.Advanced Research Center for Oral and Craniofacial SciencesOkayama University Dental School/Graduate School of Medicine, Dentistry and Pharmaceutical SciencesOkayamaJapan
  3. 3.Department of MembraneBiochemistryOkayama University Graduate School of Medicine, Dentistry and Pharmaceutical SciencesOkayamaJapan

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