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AB65311

Cytochrome c Release Assay Kit

4

(3 Reviews)

|

(52 Publications)

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.

View Alternative Names

CYC, CYCS, Cytochrome c

2 Images
Functional Studies - Cytochrome c Release Assay Kit (AB65311)
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Lab

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) (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) (ab97040) (dilution 1 : 2000).

Bands were detected at the prediced size of 12 kDa.

Functional Studies - Cytochrome c Release Assay Kit (AB65311)
  • FuncS

PubMed

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.

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/

Key facts

Sample types

Tissue, Suspension cells, Adherent cells

Reacts with

Mouse, Rat, Human

Assay type

Direct

Assay time

3h

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Product details

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.

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What's included?

{ "values": { "100Test": { "sellingSize": "100 Test", "publicAssetCode":"ab65311-100Test", "assetComponentDetails": [ { "size":"1 x 100 µL", "name":"Anti-Mouse Cyt C Antibody", "number":"AB65311-CMP02", "productcode":"" }, { "size":"1 x 100 µL", "name":"DTT II", "number":"AB65311-CMP03", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Protease Inhibitor Cocktail I", "number":"AB65311-CMP04", "productcode":"" }, { "size":"1 x 20 mL", "name":"5X Cytosol Extraction Buffer I", "number":"AB65311-CMP01", "productcode":"" }, { "size":"1 x 10 mL", "name":"Mitochondria Extraction Buffer I", "number":"AB65311-CMP05", "productcode":"" } ] } } }
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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
-20°C
Appropriate long-term storage conditions
-20°C
Storage information
-20°C
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Supplementary information

This supplementary information is collated from multiple sources and compiled automatically.

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.

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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Product protocols

For this product, it's our understanding that no specific protocols are required. You can visit:
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Target data

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.
See full target information CYCS
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Publications (52)

Recent publications for all applications. Explore the full list and refine your search

PNAS nexus 3:pgae018 PubMed38328780

2024

p17/C18-ceramide-mediated mitophagy is an endogenous neuroprotective response in preclinical and clinical brain injury.

Applications

Unspecified application

Species

Unspecified reactive species

Eda 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 Albayram

Journal of gastrointestinal oncology 14:1669-1693 PubMed37720449

2023

Esophageal cancer stem cells reduce hypoxia-induced apoptosis by inhibiting the GRP78-perk-eIF2α-ATF4-CHOP pathway .

Applications

Unspecified application

Species

Unspecified reactive species

Ruijiang Lin,Minjie Ma,Biao Han,Ya Zheng,Yuping Wang,Yongning Zhou

Cell death discovery 9:66 PubMed36797268

2023

Elephant TP53-RETROGENE 9 induces transcription-independent apoptosis at the mitochondria.

Applications

Unspecified application

Species

Unspecified reactive species

Aidan 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 Abegglen

Pharmaceutics 14: PubMed36432697

2022

Selective Induction of Intrinsic Apoptosis in Retinoblastoma Cells by Novel Cationic Antimicrobial Dodecapeptides.

Applications

Unspecified application

Species

Unspecified reactive species

Vishnu Suresh Babu,Atish Kizhakeyil,Gagan Dudeja,Shyam S Chaurasia,Veluchami Amutha Barathi,Stephane Heymans,Navin Kumar Verma,Rajamani Lakshminarayanan,Arkasubhra Ghosh

Communications biology 5:778 PubMed35995965

2022

Label-free chemical imaging of cytochrome P450 activity by Raman microscopy.

Applications

Unspecified application

Species

Unspecified reactive species

Menglu Li,Yasunori Nawa,Seiichi Ishida,Yasunari Kanda,Satoshi Fujita,Katsumasa Fujita

Cells 11: PubMed35455952

2022

Lysosomes Dysfunction Causes Mitophagy Impairment in PBMCs of Sporadic ALS Patients.

Applications

Unspecified application

Species

Unspecified reactive species

Matteo Bordoni,Orietta Pansarasa,Eveljn Scarian,Riccardo Cristofani,Roberta Leone,Valentina Fantini,Maria Garofalo,Luca Diamanti,Stefano Bernuzzi,Stella Gagliardi,Stephana Carelli,Angelo Poletti,Cristina Cereda

Journal of experimental & clinical cancer research : CR 40:248 PubMed34364401

2021

Metabolic impairment of non-small cell lung cancers by mitochondrial HSPD1 targeting.

Applications

Unspecified application

Species

Unspecified reactive species

Beatrice 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 Ceppi

Molecular oncology 16:732-749 PubMed34133843

2021

Genome-wide analysis of DNA methylation identifies the apoptosis-related gene UQCRH as a tumor suppressor in renal cancer.

Applications

Unspecified application

Species

Unspecified reactive species

Kosuke Miyakuni,Jun Nishida,Daizo Koinuma,Genta Nagae,Hiroyuki Aburatani,Kohei Miyazono,Shogo Ehata

Cell transplantation 30:9636897211024210 PubMed34105393

2021

Platelet Mitochondria Transplantation Rescues Hypoxia/Reoxygenation-Induced Mitochondrial Dysfunction and Neuronal Cell Death Involving the FUNDC2/PIP3/Akt/FOXO3a Axis.

Applications

Unspecified application

Species

Unspecified reactive species

Chun Shi,Han Guo,Xintong Liu

Molecular medicine reports 24: PubMed34036382

2021

Effects of Txk‑mediated activation of NF‑κB signaling pathway on neurological deficit and oxidative stress after ischemia‑reperfusion in rats.

Applications

Unspecified application

Species

Unspecified reactive species

Qian-Lan Xu,Jie Wu
View all publications
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We are committed to supporting your work with high-quality reagents, and we're here for you every step of the way. In the unlikely event that one of our products does not perform as expected, you're protected by our Product Promise.
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