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Cytochrome c Release Assay Kit ab65311 provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis.

Sample types

Tissue, Suspension cells, Adherent cells

Assay type

Direct

Reactive species

Mouse, Rat, Human

Assay time

3h

Images

Target data

What's included?

100 Test
Components
5X Cytosol Extraction Buffer I
1 x 20 mL
Anti-Mouse Cyt C Antibody
1 x 100 µL
DTT II
1 x 100 µL
Mitochondria Extraction Buffer I
1 x 10 mL
Protease Inhibitor Cocktail I
1 x 1 Vial

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Cytochrome c Release Assay Kit ab65311 provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis.

Key facts

Sample types

Tissue, Suspension cells, Adherent cells

Assay type

Direct

Reactive species

Mouse, Rat, Human

Assay time

3h

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

-20°C

Appropriate long-term storage conditions

-20°C

Storage information

-20°C

Notes

Cytochrome c Release Assay Kit ab65311 provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis.

The kit provides reagents to isolate a highly enriched mitochondria fraction from cytosol. The procedure is simple and easy to perform; no ultracentrifugation is required and no toxic chemicals are involved.

Cytochrome c release from mitochondria into the cytosol is determined by Western blotting using the cytochrome c antibody provided in the kit.

The anti-Cytochrome c antibody is a mouse monoclonal antibody that reacts with denatured human, mouse, and rat cytochrome c.

Cytochrome c release assay protocol summary:
- collect cells, centrifuge and wash with PBS
- resuspend in cytosol extraction buffer mix
- homogenize cells with a dounce tissue grinder
- centrifuge homogenate at 700 x g for 10 min
- collect supernatant and centrifuge at 10,000 g for 30 min, collect supernatant as cytosolic fraction
- resuspend pellet in mitochondrial extraction buffer mix and save as mitochondrial fraction
- analyze cytosolic and mitochondrial fractions in western blotting with cytochrome c antibody

This kit was previously called Cytochrome c Releasing Apoptosis Assay Kit. Cytochrome c plays an important role in apoptosis. The protein is located in the space between the inner and outer mitochondrial membranes. An apoptotic stimulus triggers the release of cytochrome c from the mitochondria into cytosol where it binds to Apaf-1. The cytochrome c/Apaf-1 complex activates caspase-9, which then activates caspase-3 and other downstream caspases. **Other apoptosis assays** For more apoptosis assays, review the .

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Product protocols

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