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  • CardioExcyte 96 SOL:用光遗传的手段起搏心肌细胞

    CardioExcyte 96 SOL:用光遗传的手段起搏心肌细胞

  • SURFE²R 96SE: 非标记高通量转运体筛选

    SURFE²R 96SE: 非标记高通量转运体筛选

  • 脂双层记录: Orbit产品系列

    脂双层记录: Orbit产品系列

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SyncroPatch 384/768PE

SyncroPatch 384/768PE

Patchliner

Patchliner

Port-a-Patch

Port-a-Patch

CardioExcyte 96

CardioExcyte 96

SURFE²R 96SE

SURFE²R 96SE

SURFE²R N1

SURFE²R N1

Orbit 16

Orbit 16

Orbit Mini

Orbit Mini

Vesicle Prep Pro

Vesicle Prep Pro

2017 - Organelle membrane derived patches: reshaping classical methods for new targets

icon pap   Port-a-Patch and   icon vpp   Vesicle Prep Pro publication in Nature Scientific Reports (2017)

Authors:
Shapovalov G., Ritaine A., Bidaux G., Slomianny, C., Borowiec A-S., Gordienko D., Bultynck G., Skryma R., Prevarskaya N.

Journal: 
Nature Scientific Reports (2017) 7:14082


Abstract:

Intracellular ion channels are involved in multiple signaling processes, including such crucial ones as regulation of cellular motility and fate. With 95% of the cellular membrane belonging to intracellular organelles, it is hard to overestimate the importance of intracellular ion channels. Multiple studies have been performed on these channels over the years, however, a unified approach allowing not only to characterize their activity but also to study their regulation by partner proteins, analogous to the patch clamp “golden standard”, is lacking. Here, we present a universal approach that combines the extraction of intracellular membrane fractions with the preparation of patchable substrates that allows to characterize these channels in endogenous protein environment and to study their regulation by partner proteins. We validate this method by characterizing activity of multiple intracellular ion channels localized to different organelles and by providing detailed electrophysiological characterization of the regulation of IP3R activity by endogenous Bcl-2. Thus, after synthesis and reshaping of the well-established approaches, organelle membrane derived patch clamp provides the means to assess ion channels from arbitrary cellular membranes at the single channel level.


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