AB65311

Cytochrome c Release Assay Kit

Name: Cytochrome c Release Assay Kit. Apoptosis assay. (ab65311) | Abcam
Brand: Abcam Limited
SKU: AB65311
Price: 870 USD
Availability: InStock
Rating: 4 (3 reviews)

(3 Reviews)

(57 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)

FuncS

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.

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

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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 product is manufactured by BioVision, an Abcam company and was previously called K257 Cytochrome c Releasing Apoptosis Assay Kit. K257-100 is the same size as the 100 test size of ab65311.

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 apoptosis assay and apoptosis marker guide.

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 10 mL", "name":"Mitochondria Extraction Buffer I", "number":"AB65311-CMP05", "productcode":"" }, { "size":"1 x 20 mL", "name":"5X Cytosol Extraction Buffer I", "number":"AB65311-CMP01", "productcode":"AB323986" }, { "size":"1 x 1 Vial", "name":"Protease Inhibitor Cocktail I", "number":"AB65311-CMP04", "productcode":"" }, { "size":"1 x 100 µL", "name":"DTT II", "number":"AB65311-CMP03", "productcode":"" }, { "size":"1 x 100 µL", "name":"Anti-Mouse Cyt C Antibody", "number":"AB65311-CMP02", "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

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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 (57)

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

Frontiers in pharmacology 15:1436786 PubMed39411073

2024

KS18, a Mcl-1 inhibitor, improves the effectiveness of bortezomib and overcomes resistance in refractory multiple myeloma by triggering intrinsic apoptosis.

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Omar S Al-Odat,Weam Othman Elbezanti,Krishne Gowda,Sandeep K Srivastava,Shantu G Amin,Subash C Jonnalagadda,Tulin Budak-Alpdogan,Manoj K Pandey

Gut microbes 16:2316932 PubMed38356294

2024

serovar Typhimurium remodels mitochondrial dynamics of macrophages via the T3SS effector SipA to promote intracellular proliferation.

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Xingmei Liu,Yutao Liu,Xinyu Zhao,Xueping Li,Ting Yao,Ruiying Liu,Qian Wang,Qiushi Wang,Dan Li,Xintong Chen,Bin Liu,Lu Feng

PNAS nexus 3:pgae018 PubMed38328780

2024

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

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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

iScience 26:108080 PubMed37860693

2023

The SARS-CoV-2 protein ORF3c is a mitochondrial modulator of innate immunity.

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Species

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Hazel Stewart,Yongxu Lu,Sarah O'Keefe,Anusha Valpadashi,Luis Daniel Cruz-Zaragoza,Hendrik A Michel,Samantha K Nguyen,George W Carnell,Nina Lukhovitskaya,Rachel Milligan,Yasmin Adewusi,Irwin Jungreis,Valeria Lulla,David A Matthews,Stephen High,Peter Rehling,Edward Emmott,Jonathan L Heeney,Andrew D Davidson,James R Edgar,Geoffrey L Smith,Andrew E Firth

The EMBO journal 42:e113118 PubMed37646198

2023

ER stress induces caspase-2-tBID-GSDME-dependent cell death in neurons lytically infected with herpes simplex virus type 2.

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Species

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Fanghui Ren,Ryo Narita,Ahmad S Rashidi,Stefanie Fruhwürth,Zongliang Gao,Rasmus O Bak,Martin K Thomsen,Georges Mgm Verjans,Line S Reinert,Søren R Paludan

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 .

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

Molecular cancer therapeutics 22:844-858 PubMed37127876

2023

Tyrosine Kinase Inhibitors Diminish Renal Neoplasms in a Tuberous Sclerosis Model Via Induction of Apoptosis.

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Uchenna Unachukwu,Jarrod Sonett,Denzel Woode,Takayuki Shiomi,Kiran Chada,Jeanine M D'Armiento

Cell death discovery 9:66 PubMed36797268

2023

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

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Species

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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.

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Species

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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.

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Unspecified application

Species

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

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Product promise

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.

For full details, please see our Terms & Conditions

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

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Cytochrome c Release Assay Kit

Functional Studies - Cytochrome c Release Assay Kit (AB65311)

Functional Studies - Cytochrome c Release Assay Kit (AB65311)

Key facts

Sample types

Reacts with

Assay type

Assay time

Complementary Products

Primary Antibodies

Secondary Antibodies

Primary Antibodies

Primary Antibodies

Product details

What's included?

Properties and storage information

Shipped at conditions

Appropriate short-term storage conditions

Appropriate long-term storage conditions

Storage information

Supplementary information

Biological function summary

Pathways

Product protocols

Target data

Publications (57)

Product promise