JoVE Logo

登录

需要订阅 JoVE 才能查看此.

Measurement of Multiple Mitochondrial Parameters in Neurons using Flow Cytometry

-- views • 1:18 min

成績單

Take fixed and permeabilized neurons in a tube.

Add a blocking buffer to prevent non-specific antibody binding.

Centrifuge the suspension and remove the supernatant. Then, resuspend the neurons in buffer.

Transfer the suspension equally into tubes containing untagged and fluorophore-tagged primary antibodies.

Incubate, allowing the antibodies to specifically bind to mitochondrial respiratory chain or MRC complex subunits, a mitochondrial DNA-associated protein, and a mitochondrial outer membrane protein within the neurons.

Remove any unbound antibodies.

Incubate with fluorophore-tagged secondary antibodies that bind to the untagged primary antibodies.

Centrifuge the mixture and discard the supernatant.

Resuspend the neurons in buffer and transfer them to microcentrifuge tubes.

Perform flow cytometry to detect the fluorescent signals on the labeled neurons, corresponding to the expression levels of MRC complex subunits and the measurement of relative mitochondrial DNA levels.

article

00:52

Measurement of Multiple Mitochondrial Parameters in Neurons using Flow Cytometry

相关视频

17 Views

JoVE Logo

政策

使用条款

隐私

科研

教育

关于 JoVE

版权所属 © 2025 MyJoVE 公司版权所有,本公司不涉及任何医疗业务和医疗服务。