JC-1 Mitochondrial Membrane

Potential Detection Kit.

Packing specification

Product Numbers: JC0050,JC0100, JC0200,

Specification: 50 times，100 times, 2 * 100 times

Storage conditions

Store at 4 ℃ away from light, avoid repeated freezing and thawing, valid for one year.

Introduction

The loss of mitochondrial membrane potential (DY) is a hallmark for apoptosis. The JC-1 Assay Kit measures the mitochondrial membrane potential in cells. In non-apoptotic cells, JC-1 (5,5’,6,6’-tetrachloro- 1,1’,3,3’-tetraethylbenzimidazolylcarbocyanine iodide) exists as a monomer in the cytosol (green) and also accumulates as aggregates in the mitochondria which stain red. Whereas, in apoptotic and necrotic cells, JC-1 exists in monomeric form and stains the cytosol green.

Background

The mitochondrial permeability transition is an important step in the induction of cellular apoptosis. During this process, the electrochemical gradient (referred to as DY) across the mitochondrial membrane collapses. The collapse is thought to occur through the formation of pores in the mitochondria by dimerized Bax or activated Bid, Bak, or Bad proteins. Activation of these pro-apoptotic proteins is accompanied by the release of cytochrome c into the cytoplasm^(1-4). The JC-1 Assay Kit uses a unique cationic dye (5,5’,6,6’-tetrachloro-1,1’,3,3’tetraethylbenzimidazolylcarbocyanine iodide) to signal the loss of the mitochondrial membrane potential⁽⁵⁾. In healthy cells, the dye stains the mitochondria bright red ⁽⁶⁾. The negative charge established by the intact mitochondrial membrane potential allows the lipophilic dye, bearing a delocalized positive charge, to enter the mitochondrial matrix where it accumulates. When the critical concentration is exceeded, J-aggregates form which become fluorescent red. In apoptotic cells, the mitochondrial membrane potential collapses, and the JC-1 cannot accumulate within the mitochondria. In these cells JC-1 remains in the cytoplasm in a green fluorescent monomeric form. Apoptotic cells, showing primarily green fluorescence, are easily differentiated from healthy cells which show red and green fluorescence. The aggregate red form has absorption/emission maxima of 585/590 nm ⁽⁵⁾. The green monomeric form has absorption/ emission maxima of 510/527 nm. Both apoptotic and healthy cells can be visualized simultaneously by fluorescence microscopy using a wide band-pass filter suitable for detection of fluorescein and rhodamine emission spectra. The JC-1 reagent is easy to use. Simply dilute the reagent in cell culture medium and add to the cells. After a 15 minute incubation, wash the cells and analyze by flow cytometry or fluorescence microscopy or fluorescence plate reader.

Warnings and Precautions

1.For Research Use Only. Not for use in diagnostic procedures.

2.Gloves, protective clothing and eyewear should be worn and safe laboratory practices followed.

Kit Components: