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Caspase Protocols in Mice

  • Varsha Kaushal
  • Christian Herzog
  • Randy S. Haun
  • Gur P. Kaushal
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1133)

Abstract

Members of the caspase family of proteases are evolutionarily conserved cysteine proteases that play a crucial role as the central executioners of the apoptotic pathway. Since the discovery of caspases, many methods have been developed to detect their activation and are widely used in basic and clinical studies. In a mouse tissue, caspase activation can be monitored by cleavage of caspase-specific synthetic substrates and by detecting cleaved caspase by western blot analysis of the tissue extract. In tissue sections, active caspase can be detected by immunostaining using specific antibodies to the active caspase. In addition, among the myriads of caspase-specific substrates known so far, cleaved fragments produced by caspases from the substrates such as PARP, lamin A, and cytokeratin-18 can be monitored in tissue sections by immunostaining as well as western blots of tissue extracts. In general, more than one method should be used to ascertain detection of activation of caspases in a mouse tissue.

Key words

Caspases Immunoblot Immunostaining Antibodies Tetrapeptide substrates Chromophore Homogenization Deparaffinization 

Notes

Acknowledgement

This work was supported by NIH grant R01 DK081690 and VA Merit Award to G.P.K. and VA Merit Award to R.S.H.

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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Varsha Kaushal
    • 1
  • Christian Herzog
    • 2
  • Randy S. Haun
    • 3
    • 4
  • Gur P. Kaushal
    • 3
    • 2
  1. 1.Biology DepartmentHendrix CollegeConwayUSA
  2. 2.Department of Internal MedicineUniversity of Arkansas for Medical SciencesLittle RockUSA
  3. 3.Central Arkansas Veterans Healthcare SystemLittle RockUSA
  4. 4.Department of Pharmaceutical SciencesUniversity of Arkansas for Medical SciencesLittle RockUSA

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