Cytochrome c Release Assay试剂盒(ab65311)
Key features and details
- Assay type: Direct
- Assay time: 3 hr
- Sample type: Adherent cells, Suspension cells, Tissue
概述
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产品名称
Cytochrome c Release Assay试剂盒 -
样品类型
Tissue, Adherent cells, Suspension cells -
检测类型
Direct -
检测时间
3h 00m -
种属反应性
与反应: Mouse, Rat, Human -
产品概述
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 -
说明
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.
性能
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存放说明
Store at -20°C. Please refer to protocols. -
组件 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 -
研究领域
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相关性
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. -
细胞定位
Mitochondrial -
数据库链接
- Entrez Gene: 54205 Human
- Omim: 123970 Human
- SwissProt: P99999 Human
相关产品
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Related Products
图片
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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.
数据表及文件
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SDS download
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Datasheet download
文献 (48)
ab65311 被引用在 48 文献中.
- 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