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Mitochondrial Transmembrane Potential Apoptosis Detection Kit (ab65310)

Submit a review Q&A (4)References (5)

Key features and details

  • Assay type: Quantitative
  • Detection method: Flow cytometry-fluorescent
  • Platform: Flow cytometer, Fluorescence microscope
  • Assay time: 30 min
  • Sample type: Adherent cells, Suspension cells

製品の概要

  • 製品名

    Mitochondrial Transmembrane Potential Apoptosis Detection Kit
    Mitochondrial Membrane Potential キット 製品一覧

  • 検出方法

    Flow cytometry-fluorescent

  • サンプルの種類

    Adherent cells, Suspension cells

  • アッセイタイプ

    Quantitative

  • 全工程の試験時間

    0h 30m

  • ステップ

    One step assay

  • 製品の概要

    This Mitochondrial Transmembrane Potential Apoptosis Detection Kit ab65310 (previously called MitoCapture Mitochondrial Apoptosis Detection Fluorometric Kit K250) provides a simple, fluorescent-based method for distinguishing between healthy and apoptotic cells by detecting the changes in the mitochondrial transmembrane potential. The kit utilizes a cationic dye that fluoresces differently in healthy vs apoptotic cells. In healthy cells, the cationic dye accumulates and aggregates in the mitocondria, giving off a bright red fluorescence. In apoptotic cells, the cationic dye cannot aggregate in the mitochondria due to the altered mitochondrial transmembrane potential, and thus it remains in the cytoplasm in its monomer form, fluorescing green. The fluorescent signals can be easily detected by fluorescence microscopy using a band-pass filter (detects FITC and rhodamine) or analyzed by flow cytometry using FITC channel for green monomers (Ex/Em = 488/530+ 30 nm) and (optional) PI channel for red aggregates (Em = 488/590+ 42 nm).


     

  • 特記事項

    Disruption of the mitochondrial transmembrane potential is one of the earliest intracellular events that occur following induction of apoptosis.

  • 試験プラットフォーム

    Flow cytometer, Fluorescence microscope

製品の特性

参考文献 (5)

ab65310 を使用した論文を発表された方は、こちらまでお知らせください。データシートに掲載させていただきます。

ab65310 は 5 報の論文で使用されています。

  • Yang G  et al. Kaempferol exhibits a synergistic effect with doxorubicin to inhibit proliferation, migration, and invasion of liver cancer. Oncol Rep 45:N/A (2021). PubMed: 33649865
  • Riera Leal A  et al. 17ß-estradiol-induced mitochondrial dysfunction and Warburg effect in cervical cancer cells allow cell survival under metabolic stress. Int J Oncol 56:33-46 (2020). PubMed: 31746421
  • Plataki M  et al. Mitochondrial Dysfunction in Aged Macrophages and Lung during Primary Streptococcus pneumoniae Infection is Improved with Pirfenidone. Sci Rep 9:971 (2019). PubMed: 30700745
  • Lin P  et al. Viral Nonstructural Protein 1 Induces Mitochondrion-Mediated Apoptosis in Mink Enteritis Virus Infection. J Virol 93:N/A (2019). PubMed: 31484746
  • Stander XX  et al. Synergistic anticancer potential of dichloroacetate and estradiol analogue exerting their effect via ROS-JNK-Bcl-2-mediated signalling pathways. Cell Physiol Biochem 35:1499-526 (2015). PubMed: 25791820

レビューと Q&A

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1-4 of 4 Abreviews or Q&A

Question

Thank you for your confirmation.
Our customer still has doubt about the differences between ab65309 and ab65310.
Because she thinks there should be some differences that's why abcam sells the same product name with different cat#.
If there's aquality issue, then she would like to purchase a kit which gives a better results.
I have explained about test size differences, but if you have more information, let me know.

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Answer

Thank you for your reply.


Please assure your customer that the two are indeed the same product. Once we have sold the remaining units of this kit under ab65309, all stock will be listed under ab65310, with the option to purchase the 25 or 100 test size.


I hope this information is helpful. Please do not hesitate to contact me if you have any additional questions.

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Question

To detect the stained cells under the microscope, the protocol said [Place the cell suspension from Step 1.g on a glass slide. Cover the cells with a glass coverslip.]
Well, the suspension from that step is 1 ml !
How can I cover this big volume on the slide by the the glass cover slip !!
Does the coverslip need to cover the 1 ml completely !!!
I need more explanation and clarification to attain this step successfully.

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Answer

You are correct, 1 ml is too much, and the instructions are too vague. You can pipet just a portion of that cell suspension onto a slide, for instance 50 ul, then coverslip. That will be 50,000 cells if you start with one million. If you find that the cells are too crowded on the slide, you can dilute the cell suspension with more buffer. Please let me know if you have more questions.

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Question

I am going to have different samples to evaluate, do you recommend cells fixation for more stable sample preparation? if the answer is yes, is there any specific protocol or fixation solution you recommend to me to use?

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Answer

We assume this assay requires living cells, and that fixation before adding the dye will give unreliable results. However, we have not compared results from fixed and unfixed samples. The question is whether the fixation itself will alter the mitochondrial transmembrane potential. I am sorry we do not have an answer at this time.

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Question

Using this kit on adherent cells, which are detaching from the surface during the assay. The apoptosis treatment is a trypsin-like enzyme, and the cells are treated for 2 hours without FBS.

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Answer

Thank you very much for your call last week and for your patience while I have been in touch with the lab regarding your enquiry.

The new kit ab65310 actually contains the same reagents that were previously included in the kit ab65309. Ab65310 is just a larger size (100 reactions versus 25), so the results should be similar between these two kits. I'm not sure why this problem is only occuring with ab65310.

I apologize but the information in the troubleshooting section of the protocol is incorrect, and there is no "binding buffer" to use as a washing solution. The cells should be resuspended with the provided Incubation Buffer after incubation with the MitoCapture solution. The components of this Incubation Buffer are proprietary and we unfortunately can not disclose the composition of this reagent.

Since the cells are treated with a trypsin-like enzyme (and trypsin causes cell detachment), and apoptosis can also cause the cells to detach, it may not be possible to keep all the cells adhered to the surface. The lab scientists recommended very gentle treatment of the cells during the assay, however it may be necessary to collect the cells after apoptosis induction and perform the protocol on the cells in suspension as written, if they are still coming off of the surface.

I hope that this information will be useful, but if you have any further questions or comments please let me know and I'll be happy to help.

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