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Antibodies & Assays for Mitochondrial Detection

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Mitochonrial apoptosis pathway.
Mitochondria are important cellular organelles that maintain crucial cellular energy balance, contain key regulators of cell death processes such as apoptosis and play a significant role in cellular oxidative stress generation. Fluorescence-based assays designed to evaluate the functional status of mitochondria are emerging as useful tools to elucidate the role of mitochondrial activity in the apoptosis cascade and other cellular processes. Loss of the mitochondrial inner transmembrane potential (ΔΨm) is often, but not always, observed to be associated with the early stages of apoptosis and is also believed to have a role in caspase independent cell death.

Collapse of mitochondrial membrane potential is believed to coincide with the permeabilization of outer mitochondrial membrane, and release of Cytochrome c and other pro-apoptotic proteins into the cytosol, which then triggers the downstream events in the apoptotic cascade.

Merck provides antibodies, inhibitors, proteins and assays to help detect and study mitochondrial proteins and membrane potential changes in apoptotic cells.

Product Highlight:
MitoLight® Mitochondrial Apoptosis Detection Kit
Apoptotic cells exhibit cytoplasmic distribution of the monomeric form of the dye (green).
Fluorescence Microscopy of NIH3T3 cells incubated with MitoLight reagent. Healthy cells contain aggregate MitoLight dye (red) in mitochondria. Apoptotic cells exhibit cytoplasmic distribution of the monomeric form of the dye (green).
The MitoLight Apoptosis Detection Kit utilizes a lipophilic cation, termed as MitoLight, as a mitochondrial activity marker. MitoLight is a mitochondrial dye that stains mitochondria in living cells in a membrane potential-dependent fashion. MitoLight partitions differently in healthy cells than in apoptotic cells. Therefore, it has been possible to use a fluorescence ratioing technique to study mitochondrial membrane potentials. In healthy cells, the dye accumulates and aggregates in the mitochondria, giving off a bright red fluorescence (λem = 585-590 nm). In apoptotic cells with altered mitochondrial membrane potential, the dye in its monomeric form stays in the cytoplasm, fluorescing green (λem = 527-530 nm), providing a ready discrimination between apoptotic and nonapoptotic cells. The fluorescence can be observed by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine) or analyzed by flow cytometry using FITC channel for green monomers (Ex/Em = 488/530) and PI channel for red aggregates (Ex/Em = 488/585).

Product Highlight:
FlowCellect MitoStress Kit
Merck’s FlowCellect MitoStress Kit includes MitoSOX Red, a live-cell permeant fluorogenic dye which targets the mitochondria and reacts with superoxide radicals and fluoresces yellow/red, and Annexin V conjugated to CF647 which binds to phosphatidylserine (PS) on the surface of apoptotic cells. The simultaneous use of these reagents allows researchers to obtain information on oxidative stress and apoptosis in one simple assay via flow cytometry.
Samples which undergo a change in mitochondrial superoxide production will show an increase in yellow/red fluorescence and samples which demonstrate increased apoptosis will show an increase in red2 fluorescence.
Dot plot analysis: Jurkat cells treated with different inducers and then stained using FlowCellect MitoStress Kit. Jurkat cells were untreated (Plot A), or treated with 150µM Antimycin A, a known inhibitor of Mitochondrial complex III which increases mitochondrial ROS production (Plot B) or with 1µM Staurosporine (Plot C) and then stained using FlowCellect MitoStress Kit. Samples which undergo a change in mitochondrial superoxide production will show an increase in yellow/red fluorescence and samples which demonstrate increased apoptosis will show an increase in red2 fluorescence.

Product Highlight:
guava® Mitochondrial Depolarization Kit
The guava Mitochondrial Depolarization Assay for the guava easyCyte Plus System allows you to monitor the loss of mitochondrial membrane potential through the use of the fluorescent cationic dye, JC-1. An optional dead cell dye, 7-AAD, is also included for discerning apoptotic cells from dead cells.

In healthy, non-apoptotic cells, the JC-1 dye enters the mitochondrial matrix, accumulates as aggregates and stains the mitochondria bright red. In apoptotic cells, the mitochondrial membrane potential collapses, and the JC-1 enters the cytoplasm as a monomeric form where it fluoresces green. Results are automatically calculated and presented in a statistical format that is easy to interpret, with total cell counts and percentages for those cell populations that are either healthy, apoptotic, or dead. For use with easyCyte Mini, easyCyte Plus.
Guava mitochondrial depolarization kit.
The pattern of JC-1 staining may vary between cell type and cell line. In this example, healthy, non-induced Jurkat cells are ~81 % polarized/healthy, ~17 % depolarized/apoptotic, and <3 % dead.

Merck's Family of Mitochondrial Detection Assays & Antibodies

DescriptionCatalog No.Assays
MitoLight Mitochondrial Apoptosis Detection Kit APT142 25 assays
MitoLight Mitochondrial Apoptosis Detection Kit APT242 100 assays
MTT Cell Growth Assay Kit CT01 5000 assays
FlowCellect MitoStress Kit FCCH100109 100 assays
FlowCellect MitoLive Kit FCCH100107 100 assays
FlowCellect MitoPotential Red Kit FCCH100105 100 assays
FlowCellect MitoDamage Kit FCCH100106 100 assays
Guava Mitochondrial Depolarization Kit 4500-0250 100 assays
Mitochondria/Cytosol Fractionation Kit MIT1000 1 Kit
Anti-Mitochondria, surface of intact mitochondria, clone 113-1 MAB1273 100 µL
Anti-Cytochrome C, clone C-7 05-479 200 µL
Anti-Cytochrome C, clone EP1326Y, Rabbit Monoclonal 04-1043 100 µL
Anti-Cytochrome c Mouse mAb (7H8.2C12) AP1029-100UG 100 µg
Anti-Cytochrome c Mouse mAb (6H2.B4) AP1030-50UG 50 µg