Cytochrome c Release Assay Kit ab65311 provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis.
-Simple and easy assay: No ultracentrifugation is required
- Includes method to separate mitochondrial and cytosolic fractions, cytochrome c is then detected by Western blot
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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- PNAS nexus 3:pgae0182024p17/C18-ceramide-mediated mitophagy is an endogenous neuroprotective response in preclinical and clinical brain injury.Applications:Unspecified applicationReactive species:Unspecified reactive speciesEda Karakaya,Natalia Oleinik,Jazlyn Edwards,Jensen Tomberlin,Randy Bent Barker,Burak Berber,Maria Ericsson,Habeeb Alsudani,Adviye Ergul,Semir Beyaz,John J Lemasters,Besim Ogretmen,Onder AlbayramPubMed 38328780
- Journal of gastrointestinal oncology 14:1669-16932023Esophageal cancer stem cells reduce hypoxia-induced apoptosis by inhibiting the GRP78-perk-eIF2α-ATF4-CHOP pathway .Applications:Unspecified applicationReactive species:Unspecified reactive speciesRuijiang Lin,Minjie Ma,Biao Han,Ya Zheng,Yuping Wang,Yongning ZhouPubMed 37720449
- Cell death discovery 9:662023Elephant TP53-RETROGENE 9 induces transcription-independent apoptosis at the mitochondria.Applications:Unspecified applicationReactive species:Unspecified reactive speciesAidan J Preston,Aaron Rogers,Miranda Sharp,Gareth Mitchell,Cristhian Toruno,Brayden B Barney,Lauren N Donovan,Journey Bly,Ryan Kennington,Emily Payne,Anthony Iovino,Gabriela Furukawa,Rosann Robinson,Bahar Shamloo,Matthew Buccilli,Rachel Anders,Sarah Eckstein,Elizabeth A Fedak,Tanner Wright,Carlo C Maley,Wendy K Kiso,Dennis Schmitt,David Malkin,Joshua D Schiffman,Lisa M AbegglenPubMed 36797268
- Pharmaceutics 14:2022Selective Induction of Intrinsic Apoptosis in Retinoblastoma Cells by Novel Cationic Antimicrobial Dodecapeptides.Applications:Unspecified applicationReactive species:Unspecified reactive speciesVishnu Suresh Babu,Atish Kizhakeyil,Gagan Dudeja,Shyam S Chaurasia,Veluchami Amutha Barathi,Stephane Heymans,Navin Kumar Verma,Rajamani Lakshminarayanan,Arkasubhra GhoshPubMed 36432697
- Communications biology 5:7782022Label-free chemical imaging of cytochrome P450 activity by Raman microscopy.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMenglu Li,Yasunori Nawa,Seiichi Ishida,Yasunari Kanda,Satoshi Fujita,Katsumasa FujitaPubMed 35995965
- Cells 11:2022Lysosomes Dysfunction Causes Mitophagy Impairment in PBMCs of Sporadic ALS Patients.Applications:Unspecified applicationReactive species:Unspecified reactive speciesMatteo Bordoni,Orietta Pansarasa,Eveljn Scarian,Riccardo Cristofani,Roberta Leone,Valentina Fantini,Maria Garofalo,Luca Diamanti,Stefano Bernuzzi,Stella Gagliardi,Stephana Carelli,Angelo Poletti,Cristina CeredaPubMed 35455952
- Journal of experimental & clinical cancer research : CR 40:2482021Metabolic impairment of non-small cell lung cancers by mitochondrial HSPD1 targeting.Applications:Unspecified applicationReactive species:Unspecified reactive speciesBeatrice Parma,Vignesh Ramesh,Paradesi Naidu Gollavilli,Aarif Siddiqui,Luisa Pinna,Annemarie Schwab,Sabine Marschall,Shuman Zhang,Christian Pilarsky,Francesca Napoli,Marco Volante,Sophia Urbanczyk,Dirk Mielenz,Henrik Daa Schrøder,Marc Stemmler,Heiko Wurdak,Paolo CeppiPubMed 34364401
- Molecular oncology 16:732-7492021Genome-wide analysis of DNA methylation identifies the apoptosis-related gene UQCRH as a tumor suppressor in renal cancer.Applications:Unspecified applicationReactive species:Unspecified reactive speciesKosuke Miyakuni,Jun Nishida,Daizo Koinuma,Genta Nagae,Hiroyuki Aburatani,Kohei Miyazono,Shogo EhataPubMed 34133843
- Cell transplantation 30:96368972110242102021Platelet Mitochondria Transplantation Rescues Hypoxia/Reoxygenation-Induced Mitochondrial Dysfunction and Neuronal Cell Death Involving the FUNDC2/PIP3/Akt/FOXO3a Axis.Applications:Unspecified applicationReactive species:Unspecified reactive speciesChun Shi,Han Guo,Xintong LiuPubMed 34105393
- Molecular medicine reports 24:2021Effects of Txk‑mediated activation of NF‑κB signaling pathway on neurological deficit and oxidative stress after ischemia‑reperfusion in rats.Applications:Unspecified applicationReactive species:Unspecified reactive speciesQian-Lan Xu,Jie WuPubMed 34036382
Key facts
- Sample types
- Tissue, Suspension cells, Adherent cells
- Assay type
- Direct
- Reactive species
- Mouse, Rat, Human
- Assay time
- 3h
Target data
Function
Electron carrier protein. The oxidized form of the cytochrome c heme group can accept an electron from the heme group of the cytochrome c1 subunit of cytochrome reductase. Cytochrome c then transfers this electron to the cytochrome oxidase complex, the final protein carrier in the mitochondrial electron-transport chain. Plays a role in apoptosis. Suppression of the anti-apoptotic members or activation of the pro-apoptotic members of the Bcl-2 family leads to altered mitochondrial membrane permeability resulting in release of cytochrome c into the cytosol. Binding of cytochrome c to Apaf-1 triggers the activation of caspase-9, which then accelerates apoptosis by activating other caspases.
Alternative names
CYC, CYCS, Cytochrome c
What's included?
Recommended products
Cytochrome c Release Assay Kit ab65311 provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis.
-Simple and easy assay: No ultracentrifugation is required
- Includes method to separate mitochondrial and cytosolic fractions, cytochrome c is then detected by Western blot
- Individual kit components also available for purchase with a minimum order of 20 units. Contact us to discuss your needs.
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 .
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the Support and downloads section.
Supplementary info
- This supplementary information is collated from multiple sources and compiled automatically.
- Activity summary
Cytochrome c also known as cyt c is a small heme protein with a mass of approximately 12 kDa. It is expressed in the mitochondria of eukaryotic cells across different species. Its mechanical role involves electron transfer within the mitochondrial electron transport chain. Cytochrome c transfers electrons between complex III (cytochrome bc1 complex) and complex IV (cytochrome c oxidase complex) playing a critical role in cellular respiration and energy production.
- Biological function summary
Cytochrome c participates in both energy generation and apoptosis. It functions as part of the electron transport chain complex contributing to the maintenance of the mitochondrial membrane potential. When released into the cytosol cytochrome c initiates the apoptotic cascade by binding to Apaf-1 which then forms the apoptosome complex along with procaspase-9. This triggers subsequent caspase activation leading to programmed cell death.
- Pathways
Cytochrome c plays a central role in the intrinsic pathway of apoptosis and oxidative phosphorylation. In the apoptotic pathway after release from mitochondria it interacts with Apaf-1 and procaspase-9 eventually activating caspase-3 and other executioner caspases. Additionally during oxidative phosphorylation cytochrome c works closely with cytochrome bc1 and cytochrome c oxidase in the electron transport chain facilitating ATP synthesis.
- Associated diseases and disorders
Cytochrome c is linked with conditions such as cancer and neurodegenerative diseases. Its release from mitochondria is associated with apoptotic cell death which can either suppress tumor development or contribute to cancer cell resistance. In neurodegenerative disorders like Alzheimer's disease impaired cytochrome c function can result in cellular energy deficits and increased apoptosis. The interaction with Apaf-1 is vital in these processes highlighting the importance of cytochrome c in pathology.
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2 product images
Functional Studies - Cytochrome c Release Assay Kit (ab65311)
5x10e7 Jurkat cells were cultured in the absence (1-2) or presence of 2 uM Camptothecin (CPT) (Camptothecin, DNA topoisomerase inhibitor ab120115) for 24 hours (3-4) or with 10 uM CPT for 4 hours (5-6). 30 uL cytosolic (1, 3, 5) and mitochondrial (2, 4, 6) extracts were loaded onto the gel. Membranes were probed with anti-Cytochrome C Mouse MAb (ab65311) (dilution 1:200) followed by Goat Anti-Mouse IgG (HRP) (Goat Anti-Mouse IgG H&L (HRP) preadsorbed ab97040) (dilution 1:2000).
Bands were detected at the prediced size of 12 kDa.
Sun Q., PLoS One 9(6), Fig 4. doi: 10.1371/journal.pone.0098866. Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/Functional Studies - Cytochrome c Release Assay Kit (ab65311)
Inhibition of cytochrome c release from mitochondria in SK-N-SH cells.
Cytochrome C release was measured using Cytochrome C releasing apoptosis assay kit (ab65311). Blots showing immunoreactive bands for cytochrome c in cytosol (Image A). Data was expressed in fold-increase of cytochrome c compared to vehicle. Protein expression levels were normalized to β-actin (Figure B). Blots (Image C) of immunoreactive bands for cytochrome C in mitochondria. Figure D shows a fold-increase of cytochrome C compared to vehicle. Protein expression levels were normalized to COX IV.
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