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Product Details
| Cat # +Size | K257-100 |
|---|---|
| Size | 100 assays |
| Detection Method | Western blotting |
| Species Reactivity | Mammalian |
| Applications | Effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis. The kit provides an easy-to-use procedure and reagents for separating a mitochondria-enriched fraction from cytosol. |
| Features & Benefits | • Simple procedure; takes ~3 hours • Fast and convenient • Cytochrome c releasing from mitochondria into cytosol is then determined by Western blotting using the cytochrome c antibody provided in the kit. The procedure is simple and straightforward. No ultracentrifugation is required. |
| Kit Components | • Mitochondria Extraction Buffer • 5X Cytosol Extraction Buffer • DTT (1 M) • 500X Protease Inhibitor Cocktail • Anti-Cytochrome c mouse mAb (0.2 mg/ml) |
| Storage Conditions | -20°C |
| Shipping Conditions | Gel Pack |
| USAGE | For Research Use Only! Not For Use in Humans. |
Details
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. BioVision’s Cytochrome c Releasing Apoptosis Assay Kit provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis. The kit provides unique formulations of reagents to isolate a highly enriched mitochondria fraction from cytosol. The procedure is so simple and easy to perform, no ultracentrifugation is required and no toxic chemicals are involved. Cytochrome c releasing from mitochondria into cytosol is then determined by Western blotting using the cytochrome c antibody provided in the kit.
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Do we have to perform a denaturation step of the samples by heating at 95°C during 5 min as usual?
Yes, you heat with the loading buffer as you would for a western blot using SDS-PAGE
Are the Cytosolic Fraction and Mitochondrial Fraction suitable for a BCA assay?
Yes, a BCA assay can be used but it is beter to use a reducing agent compatible one: http://www.biovision.com/bca-protein-assay-kit-reducing-agent-compatible-6891.html
Can the kit work on bacteria or yeast cells?
The kit has been standardized for mammalian cells only
What is the exact volume of sample required for this assay?
There is no specific volume we can recommend for the amount any sample to be used since it is completely sample concentration and quality based. You have to do a pilot expt with multiple sample volumes to determine the optimal volume which gives a reading within the linear range of the standard curve. Please refer to the citations for this product to see what other clients have used with similar sample types.
Do you have trial sizes of this kit?
Unfortunately, we do not have trial sizes of this kit available. However, if you are based in the US or Canada, we will give you a 10% off list price introductory discount on its purchase price. If you are based out of this area please contact yopur regional BioVision distributor.
What is the shelf life of this kit?
BioVisions’s assay kits expire 6 months from the date of shipment. Some components of the kit may expire sooner as mentioned in the data sheet
Can we purchase individual components of this kit?
Yes, you can purchase any of the kit's components without the whole kit. Please refer to the component Cat #s mentioned on the datasheet for ordering.
Can we use an alternate buffer for sample preparation (cell lysis, sample dilutions etc)?
Our assay buffers are optimized for the reactions they are designed for. They not only contain some detergents for efficient lysis of your cells/tissue, but also contain some proprietary components required for the further reactions. Therefore, we highly recommend using the buffers provided in the kit for the best results.
Huang-Ju Tu. The anticancer effects of MPT0G211, a novel HDAC6 inhibitor, combined with chemotherapeutic agents in human acute leukemia cells. Clin Epigenetics, Dec 2018; 30594243.
Malapelle, Umberto et al. (2017) USP7 inhibitors, downregulating CCDC6, sensitize lung neuroendocrine cancer cells to PARP-inhibitor drugs, Lung Cancer. 2017 May;107:41-49.
Shyam, Hari et al. (2017) Centchroman induces redox-dependent apoptosis and cell-cycle arrest in human endometrial cancer cells, Apoptosis. 2017 Apr;22(4):570-584.
Giorgi et al., p53 at the endoplasmic reticulum regulates apoptosis in a Ca2+-dependent manner. PNAS, Feb 2015; 112: 1779 - 1784.
For more citations of this product click here
