Titlebook: BCL-2 Family Proteins; Methods and Protocol Evripidis Gavathiotis Book 2019 Springer Science+Business Media, LLC, part of Springer Nature 2

Sinus-Node

Leo Man-Lit Cheang,Catherine McBridectivation and apoptotic cell death. To generate pores, BAK and BAX undergo major changes including BAX translocation to the outer membrane, and partial unfolding, dimerization, and oligomerization. Here we describe biochemical protocols that can be used on most cell types to gain a population overview of BAK and BAX status.

Initiative

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Probing BAK and BAX Activation and Pore Assembly with Cytochrome , Release, Limited Proteolysis, anctivation and apoptotic cell death. To generate pores, BAK and BAX undergo major changes including BAX translocation to the outer membrane, and partial unfolding, dimerization, and oligomerization. Here we describe biochemical protocols that can be used on most cell types to gain a population overview of BAK and BAX status.

Yag-Capsulotomy

Fracture

抗生素

Chivalrous

Assessment of Dynamic BCL-2 Protein Shuttling Between Outer Mitochondrial Membrane and Cytosol,nd shows correlation between relative BAX and BAK localizations and cellular predisposition to apoptosis. The retrotranslocation of BCL-2 proteins from the OMM can be measured using fluorescence-labeled protein in intact cells or endogenous protein from isolated heavy membrane fractions.

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Quantification of BCL-2 Family Members by Flow Cytometry,blotting. Here we describe a robust and reproducible method to measure the expression levels of the BCL-2 family members, BCL-2, BCL-X., and MCL-1 by quantitative flow cytometry (QFCM) using validated antibodies.

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Methods to Probe Conformational Activation and Mitochondrial Activity of Proapoptotic BAK,apoptosis, BAK undergoes conformational changes to form the active species associated with apoptotic pores. We describe methods to purify mitochondria for MOMP assays and to detect conformational changes in native BAK associated with MOMP by using limited proteolysis and cross-linking analyses.