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.
The Safety Datasheet for this product has been updated for certain countries. Please check the current version in the SDS download section.
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 5X Cytosol Extraction Buffer I 1 x 20ml Anti-Mouse Cyt C Antibody 1 x 100µl DTT II 1 x 100µl Mitochondria Extraction Buffer I 1 x 10ml Protease Inhibitor Cocktail I 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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Inhibition of cytochrome c release from mitochondria in SK-N-SH cellsSun 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/
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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Functional assays: Cytochrome c Releasing Apoptosis 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.
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 (52)
ab65311 は 52 報の論文で使用されています。
- Karakaya E et al. p17/C18-ceramide-mediated mitophagy is an endogenous neuroprotective response in preclinical and clinical brain injury. PNAS Nexus 3:pgae018 (2024). PubMed: 38328780
- Preston AJ et al. Elephant TP53-RETROGENE 9 induces transcription-independent apoptosis at the mitochondria. Cell Death Discov 9:66 (2023). PubMed: 36797268
- Lin R et al. Esophageal cancer stem cells reduce hypoxia-induced apoptosis by inhibiting the GRP78-perk-eIF2α-ATF4-CHOP pathway in vitro. J Gastrointest Oncol 14:1669-1693 (2023). PubMed: 37720449
- 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