Titlebook: Caspases,Paracaspases, and Metacaspases; Methods and Protocol Peter V. Bozhkov,Guy Salvesen Book 2014 Springer Science+Business Media New Y

Carcinogen

General In Vitro Caspase Assay Procedures their activation mechanisms, and the identification of their substrates were made possible by the availability of sufficient amounts of enzymatically pure caspases. The current chapter describes at length the expression, purification, and basic enzymatic characterization of apoptotic caspases.

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Global Identification of Caspase Substrates Using PROTOMAP (Protein Topography and Migration Analysin migration indicative of proteolytic processing. When applied to cells undergoing apoptosis, this unbiased global method provides a snapshot of the topography and magnitude of proteolytic events associated with programmed cell death.

REIGN

Metacaspase: An Arginine-Specific Peptidasee expression systems, metacaspase-deficient yeast cells (.) have been instrumental in studying the activity of . metacaspase (LmjMCA). Here, we describe techniques for purification of LmjMCA and its activity measurement, providing a platform for further identification of LmjMCA substrates.

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Preparation of , Seedling Proteomes for Identifying Metacaspase Substrates by N-terminal COFRADICwe describe a protocol for the preparation of plant tissue proteomes, including differential isotopic labelling allowing for a comparison of in vivo N-terminomes that serves as the starting point for N-terminal COFRADIC studies.

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Measurement of Caspase Activation in Mammalian Cell Culturesactivities of unique caspases are key determinants of the apoptotic process. This chapter describes a set of experimental protocols available for characterization, quantification and inhibition of caspase activities in mammalian cell cultures, including immunoblotting, usage of synthetic substrates, flow cytometry, and microscopic techniques.

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Robust Evacuation Planning for Urban Areas cell death execution events. If a protein is cleaved by CED-3 in vitro, this protein could be a potential CED-3 substrate in vivo. Here, we describe the method for purification of active CED-3 caspase. We will also describe in vitro assays for determining CED-3 proteolytic activity, CED-3 substrates, and CED-3 cleavage sites in the substrates.

Lethargic

Imaging Oral Biofilm and Plaque,ot full-length, Caspase-3. Although raised against human cleaved Caspase-3, the CC3 antibody cross-reacts in other species and detects cleaved caspases, most notably DrICE and Dcp-1, in .. This protocol describes the procedure for use of the CC3 antibody to detect caspase activity in larval imaginal discs in ..

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