Overview

  • Product name
    Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer)
    See all Mitochondria Isolation kits
  • Sample type
    Tissue
  • Assay type
    Quantitative
  • Product overview

    ab110169 (MS851) contains sufficient reagents for up to 10 benchtop mitochondria isolations from hard or soft tissues. Isolated mitochondria can be analyzed by Western Blotting, Immunocapture, or Activity assay.

  • Notes

    Tested tissues: brain, liver and heart

Properties

Images

  • Rat liver mitochondria were isolated using Abcam's ab110169/MS851 Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer). 2 µg (lane 1) and 10 µg (lane 2) of mitochondria were tested with Abcam's ab110413/MS604 (MitoProfile® Total OXPHOS Rodent WB Antibody Cocktail). Mitochondria are enriched and so Western blot band intensity of mitochondrial proteins is greater in these mitochondrial preparations when compared to homogenate at respective amounts, see lane 3 and 4

Protocols

References

This product has been referenced in:
  • Sun J  et al. Anthocyanins isolated from blueberry ameliorates CCl4 induced liver fibrosis by modulation of oxidative stress, inflammation and stellate cell activation in mice. Food Chem Toxicol 120:491-499 (2018). Read more (PubMed: 30056145) »
  • Villeneuve LM  et al. Proteomic and functional data sets on synaptic mitochondria from rats with genetic ablation of Parkin. Data Brief 20:568-572 (2018). Read more (PubMed: 30197913) »
See all 9 Publications for this product

Customer reviews and Q&As

1-10 of 22 Abreviews or Q&A

Question
Answer

Thank you for contacting us.

Samples may be prepared as purified mitochondria, crudemitochondria. In most cases whole tissue or cell lysates aresuitable but these may require some sample optimization. Mitochondria samples can be prepared bysimple differential centrifugation of homogenized samples. While any crude isolation may be effective, we recommend ourhttps://www.abcam.com/Mitochondria-Isolation-Kit-for-Cultured-Cells-ab110170.htmlhttps://www.abcam.com/Mitochondria-Isolation-Kit-for-Cultured-Cells-ab110170.htmlorhttps://www.abcam.com/Mitochondria-Isolation-Kit-for-Cultured-Cells-with-Dounce-Homogenizer-ab110171.html(with Dounce homogenizer) if using cultured myocytes such as HL-1 cells.If using heart tissues we would recommendhttps://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-ab110168.htmlhttps://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-ab110168.htmlorhttps://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-with-Dounce-Homogenizer-ab110169.html(with Dounce homogenizer).



This kit was optimized to extract complex I into its native form (with all 47 subunits assembled in to a large complex). The sample extraction is native so care needs to be taken to extract the samples at equivalent starting concentrations to maintain the protein: detergent ratio and maintain enzyme integrity and activity.Specifically for this product - the extraction is performed by adding 1/10 volume of the 10X Detergent to samples at 5.5mg/mL



I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.




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Answer

Please see below for further clarification regarding the following questions:

Is it OK to extract mitos from frozen tissues?

A: yes, you can extract mitos from frozen tissues but be aware that freezen/ thaw cycle would weaken the cell membrane and cause less recovery of intact mitochondria.
Are there differences between the extraction from frozen mitos versus fresh mitos?

A: yes, there will be slightly loss if you extract enzyme from frozen mitos versus fresh mitos due to the degradation. Adding protease inhibitor to your mito prep would minimize the loss.
Does the extraction time point from a frozen tissue matters?

A: No, as long as the tissues are properly frozen, the variant between different time points should be minimum (within weeks). However, multiple freezen/thaw cycle will cause difference. If you plan to extract sample from different days, make sure you make aliquots of your samples. if you plan to compare the enzyme activity from different samples, we highly recommend to prep the samples under the same condition.

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Answer

Thank you for contacting us. Yes, we will guarantee ab110169 for use on any mammalian tissues. While it would not qualify for our Abtrial discount program, it will be covered under our Abpromise guarantee. If you have any trouble using this kit on human samples, please let us know within six months of purchase and we will be happy to offer you a free of charge replacement, credit, or refund.

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Answer

Thank you for your enquiry regarding ab110169: Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer)

It is not clear at which step the samples got bacterial contaminated.

Unfortunately, it is difficult to remove bacteria completely without loss of mitochondria, their sizes are very similar. Genome wise, mitochondrial genome also overlapped heavily with bacterial genome. The only way is to redo the mitochondria prep and use the sample fresh.


If you need any further assistance in the future, please do not hesitate to contact me.

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Answer

Gracias por contactarnos.

Lamento mucho que el kit no haya llegado en perfecto estado a vuestro laboratorio.

¿Podrías por favor confirmarme el número de pedido del kit?

También agradecería que me enviaseis una foto del embolo roto para nuestros registros de calidad y poder trazar este asunto y detectar la fuente del problema.

De nuevo, disculpad las molestias, y espero tu respuesta.

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Answer

Thank you fro your enquiry.

I am sorry to confirm we would not recommend to use ab109718 Cell Fractionation Kit - HTor ab109719 Cell Fractionation Kit Standardfor isolation of mitochondria from tissue. They are designed for cell culture single cell suspension samples only. The issue with solid tissue is in the difficulty to prepare the starting material for the fractionation: single cell suspension without breaking the cells.

If you want to isolate mitochondria, then I can recommend to consider ab110168 or ab110169 instead. These are designed for mitochondrial isolation from tissue.

https://www.abcam.com/index.html?datasheet=110168 (or use the following: https://www.abcam.com/index.html?datasheet=110168).

https://www.abcam.com/index.html?datasheet=110169 (or use the following: https://www.abcam.com/index.html?datasheet=110169).

The important steps to consider from the protocol provided on the datasheetare as follows:

Spin 1 isolates the unbroken cells and nucleiSpin 2 isolates the mitochondria. The remaining supernatant after spin 2 contains mostly cytosolic proteins –the protocol suggests to save this supernatant sample (step 12) and these could be used.

There are wash steps to clean up the mitochondria (Steps 13-15) However the supernatant sample could also be washed by spinning at 12000g repeatedly and collecting the supernatant each time.

I hope this will be helpful to you. If you have any further questions, please do not hesitate to contact us.

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Answer


Please do not use a sonicatorin order to destroy your cells, as this will result in a complete lysis of the mitochondria as well. This means you will not be able to analyse the sample.



However, we do have a kit in which we delivers the grinder as well:

https://www.abcam.com/index.html?datasheet=110169 (or use the following: https://www.abcam.com/index.html?datasheet=110169).


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Answer

Thank you for your inquiry.

I am happy to confirm that our lab does not see any specific problems with the experimental set up and buffers your customer is planning to use.

Cardiac tissue should be an excellent materials for isolating mitochondria.

I can also confirm that the homogenizer the lab uses a has no problems in disrupting the tissue and cells, they do this routinely.

I hope I could re-assure your customer that ab110169 is the right choice for their experimentsand wish them good luck with their research.

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Answer

Thank you for contacting us.

I have attached the SDS for each kit (Mitochondria Isolation Kit for Tissue) with and without Dounce homogenizer (ab110169 and ab110168), since they are slightly different.

I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

Use our products? Submit an Abreview. Earn rewards!
https://www.abcam.com/abreviews

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1-10 of 22 Abreviews or Q&A

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