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

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Flow Cytometry-Based Detection and Analysis of BCL-2 Family Proteins and Mitochondrial Outer Membraderstanding of the mechanistic interactions between these proteins continues to evolve in normal and malignant cells. The functional variation by individual BCL-2 proteins in different cell types has driven clinical therapeutic development in targeting individual BCL-2 members with the goal of fine-

Minatory

Investigating BCL-2 Family Protein Interactions in Yeast, yeast . may look out of place. However, when grown under adequate conditions, yeast cells have mitochondria that have similar properties as those of mammalian cells, and are able to be targeted by mammalian Bcl-2 family members. Yeast thus provides a neutral cellular background to study how protein

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Liposomal Permeabilization Assay to Study the Functional Interactions of the BCL-2 Family,y BCL-2 family members have been classified in three groups based on structural homology and function. The multidomain antiapoptotic proteins promote survival, whereas the multidomain and the BH3-only proapoptotic members induce cell death. Because the interaction among the BCL-2 family members occu

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defray

Assessment of Dynamic BCL-2 Protein Shuttling Between Outer Mitochondrial Membrane and Cytosol,nd BAK but also prosurvival family members, like BCL-x. or MCL-1, translocate to the outer mitochondrial membrane (OMM) and retrotranslocate from the mitochondria back into the cytosol. The resulting equilibrium produces a broad range of localization pattern observed for BAX and BAK in human cells a

GREG

Quantification of BCL-2 Family Members by Flow Cytometry,ure the expression levels of specific proteins in a cell population of interest without the need to physically separate out the cells from within a heterogeneous population by using the appropriate cell-specific markers. It also requires fewer cells than other traditional techniques such as Western

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Methods to Probe Calcium Regulation by BCL-2 Family Members,ortant function is the release of calcium from the endoplasmic reticulum (ER), which critically contributes to the process of apoptosis. Here, we describe a protocol to measure calcium levels in the ER, mitochondria, and cytosol, with specific consideration of BCL-2 family biology.

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背書

Probing BAK and BAX Activation and Pore Assembly with Cytochrome , Release, Limited Proteolysis, an These two proteins generate pores in the mitochondrial outer membrane that release factors such as cytochrome . into the cytosol to trigger caspase activation and apoptotic cell death. To generate pores, BAK and BAX undergo major changes including BAX translocation to the outer membrane, and partia