Cytochrome c Release Assay Kit (ab65311)
Key features and details
- Assay type: Direct
- Assay time: 3 hr
- Sample type: Adherent cells, Suspension cells, Tissue
製品の概要
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製品名
Cytochrome c Release Assay Kit -
サンプルの種類
Tissue, Adherent cells, Suspension cells -
アッセイタイプ
Direct -
全工程の試験時間
3h 00m -
種交差性
交差種: Mouse, Rat, Human -
製品の概要
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 -
特記事項
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.
Abcam has not and does not intend to apply for the REACH Authorisation of customers’ uses of products that contain European Authorisation list (Annex XIV) substances.
It is the responsibility of our customers to check the necessity of application of REACH Authorisation, and any other relevant authorisations, for their intended uses.
製品の特性
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保存方法
Store at -20°C. Please refer to protocols. -
内容 100 tests 100 tests 5X Cytosol Extraction Buffer I 1 x 20ml 1 x 20ml Anti-Mouse Cyt C Antibody 1 x 100µl 1 x 100µl DTT II 1 x 110µl 1 x 110µl Mitochondria Extraction Buffer I 1 x 10ml 1 x 10ml Protease Inhibitor Cocktail I 1 vial 1 vial -
研究分野
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関連性
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. -
細胞内局在
Mitochondrial -
参照データベース
- Entrez Gene: 54205 Human
- Omim: 123970 Human
- SwissProt: P99999 Human
関連製品
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Related Products
画像
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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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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.
データシートおよび資料
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SDS download
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Datasheet download
参考文献 (48)
ab65311 は 48 報の論文で使用されています。
- Bordoni M et al. Lysosomes Dysfunction Causes Mitophagy Impairment in PBMCs of Sporadic ALS Patients. Cells 11:N/A (2022). PubMed: 35455952
- Li M et al. Label-free chemical imaging of cytochrome P450 activity by Raman microscopy. Commun Biol 5:778 (2022). PubMed: 35995965
- Suresh Babu V et al. Selective Induction of Intrinsic Apoptosis in Retinoblastoma Cells by Novel Cationic Antimicrobial Dodecapeptides. Pharmaceutics 14:N/A (2022). PubMed: 36432697
- Xu W et al. Apaf-1 Pyroptosome Senses Mitochondrial Permeability Transition. Cell Metab 33:424-436.e10 (2021). PubMed: 33308446
- Salimi A et al. Persian Gulf Snail Crude Venom (Conus textile): A Potential Source of Anti-Cancer Therapeutic Agents for Glioblastoma through Mitochondrial-Mediated Apoptosis. Asian Pac J Cancer Prev 22:49-57 (2021). PubMed: 33576212