Format

Send to

Choose Destination
See comment in PubMed Commons below
Nat Protoc. 2007;2(12):3111-4.

Assay for apoptosis using the mitochondrial probes, Rhodamine123 and 10-N-nonyl acridine orange.

Author information

1
Laboratory of Antineoplastic Pharmacology, Department of Obstetrics and Gynaecology, Catholic University of the Sacred Heart, Rome, Italy. cferlini@rm.unicatt.it

Abstract

Apoptosis plays a pivotal role in the regulation of cell turnover, and a defect or an excess of apoptosis has been implicated in several human diseases. Apoptosis is activated from an extracellular death signal, or from an internal pathway starting from the endoplasmatic reticulum or the mitochondria. To investigate the mitochondrial compartment during apoptosis, we have established a protocol using fluorochromes and flow cytometry to probe the structure and function of mitochondria kinetically. The protocol could be applied to whole cells or to isolated mitochondria. In the first case, cells are counterstained with ethidium bromide (EB) to evaluate plasma membrane function. The presence of the electrochemical gradient in the mitochondria is probed with Rhodamine123 (Rh123), whereas the structure and the integrity of mitochondria are assessed using 10-N-nonyl-acridine orange (NAO). Not considering the time requested for cell/mitochondria preparation and the activation of apoptosis, the protocol lasts <1 h.

PMID:
18079710
DOI:
10.1038/nprot.2007.397
[Indexed for MEDLINE]
PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for Nature Publishing Group
    Loading ...
    Support Center