Cytochrome c Releasing Apoptosis Assay试剂盒(ab65311)

概述

  • 产品名称Cytochrome c Releasing Apoptosis Assay试剂盒
    参阅全部 Cytochrome c 试剂盒
  • 测试
    100 x 1 assay
  • 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

    Abcam's Cytochrome c Releasing Apoptosis Assay Kit provides an effective means for detecting cytochrome c translocation from mitochondria into cytosol during apoptosis. The kit provides unique formulations of reagents to isolate a highly enriched mitochondria fraction from cytosol. The procedure is so simple and easy to perform, no ultracentrifugation is required and no toxic chemicals are involved. Cytochrome c releasing from mitochondria into cytosol is then determined by Western blotting using the cytochrome c antibody provided in the kit.
    Visit our FAQs page for tips and troubleshooting.

     

    The anti-Cytochrome c antibody is a mouse monoclonal antibody that reacts with denatured human, mouse, and rat cytochrome c.

  • 说明

    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.

  • 经测试应用适用于: WBmore details

性能

  • 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

相关产品

应用

Our Abpromise guarantee covers the use of ab65311 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

应用 Ab评论 说明
WB Use a concentration of 1 µg/ml. Predicted molecular weight: 11.7 kDa.

Cytochrome c Releasing Apoptosis Assay Kit 图像

  • 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 (ab6311) (dilution 1:200) followed by Goat Anti-Mouse IgG (HRP) (ab97040) (dilution 1:2000).

    Bands were detected at the prediced size of 12 kDa.

实验方案

Cytochrome c Releasing Apoptosis Assay Kit (ab65311)参考文献

This product has been referenced in:
  • Koontz J & Kontrogianni-Konstantopoulos A Competition through dimerization between antiapoptotic and proapoptotic HS-1-associated protein X-1 (Hax-1). J Biol Chem 289:3468-77 (2014). WB ; Rat . Read more (PubMed: 24347163) »
  • Sun Q  et al. Protective effects of astragaloside IV against amyloid beta1-42 neurotoxicity by inhibiting the mitochondrial permeability transition pore opening. PLoS One 9:e98866 (2014). WB ; Human . Read more (PubMed: 24905226) »

See all 2 Publications for this product

Product Wall

1-

Are the Cytosolic Fraction and Mitochondrial Fraction per se suitable for a BCA dosage to determine the volume necessary to obtain 10 ug? -
Yes, a BCA assay can be used but it is better to use a reducing agent compatible assay.
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The antibody from this kit is now available separately on our catalog as product ab189738 http://www.abcam.com/cytochrome-c-mouse-mab-ab189738.html.

Abreviews
Type: Tissue lysate
Amount of protein used: 35 ug
Specification: Heart
Reduce/denaturing condition: Reduced, Denaturing, 12% gel
Blocking buffer/concentration/time/temperature: BSA/5%/1 h/25oC
Antibody dilution: 1:1000
Incubation time/temperature: 18 h/4oC
Diluent: BSA
Secondary antibody information: Non-Abcam antibody/Donkey-anti-mouse/Polyclonal/HRP/1:10000
Detection method: ECL+
Exposure: 5 s
Observed band size: Specific: 12 kDa
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Abcam user community

Verified customer

提交于 Jul 05 2013

Abreviews
Type: Tissue lysate
Amount of protein used: 35 ug
Specification: Heart
Reduce/denaturing condition: Reduced, Denaturing, 12% gel
Blocking buffer/concentration/time/temperature: BSA/5%/1 h/25oC
Antibody dilution: 1:1000
Incubation time/temperature: 18 h/4oC
Diluent: BSA
Secondary antibody information: Non-Abcam antibody/Donkey-anti-mouse/Polyclonal/HRP/1:10000
Detection method: ECL+
Exposure: 5 s
Observed band size: Specific: 12 kDa
Username

Abcam user community

Verified customer

提交于 Jul 05 2013

Thank you for contacting us.

Yes, homogenization is a critical part of this protocol. You need to ensure that the cells have lysed well and released the cellular fractions in order to ensure that you see the relative difference in the levels o...

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Thank you for contacting us and your interest in our products.

As discussed over the phone I have looked further into your query in regards to the Cytochrome c Releasing Apoptosis Assay Kit (ab65311).

In reply to your questions, when ...

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Thank you for contacting us.

The lab let me know the following:

The cytosolic and the mitochondrial extraction buffers detergent based. But the exact composition is proprietary.
The nucleus will be a part of the mitochondrial ...

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Thank you for your telephone enquiry yesterday.

I can confirm that to reconstitute the protease inhibitor cocktailprovided withab65311 Cytochrome c Releasing Apoptosis Assay Kit:

Add 250 ìl DMSO to dissolve the 500X Proteas...

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Thank you for your interest in the Mitochondria/Cytosol Fractionation Kit (ab65320) and Cytochrome c Releasing Apoptosis Assay Kit (ab65311). Both kits are essentially the same in the preparation of the samples, however, following the separation of...

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Thank you for your enquiry. Theoretically this kit can be used with plant samples as long as the samples are prepared as indicated in the protocol provided with the data sheet. However, I am sorry to confirm that plant samples have not been tested ...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"