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    products/sample-preparation-kits/mitochondria-isolation-kit-for-tissue-with-dounce-homogenizer-ab110169.pdf

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Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer) (ab110169)

  • Datasheet
  • SDS
  • Protocol Booklet
Submit a review Q&A (22)References (18)

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Western blot - Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer) (ab110169)

    Key features and details

    • Assay type: Quantitative
    • Sample type: Tissue

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    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

    • Storage instructions

      Store at +4°C. Please refer to protocols.
    • Components 1 kit
      Dounce Homogenizer (2 ml size) with pestles 1 unit
      Isolation Buffer 1 x 100ml
      Wash Buffer 1 x 30ml
    • Research areas

      • Kits/ Lysates/ Other
      • Kits
      • Molecular Biology Kits
      • Cell Fractionation Kits
      • Kits/ Lysates/ Other
      • Kits
      • Extraction Kit
      • Mitochondria
    • Alternative names

      • mitochondria purification
      • mitochondrial isolation
      • mitochondrial purification

    Associated products

    • Related Products

      • Mitochondria Isolation Kit for Tissue (ab110168)
      • Mitochondria Isolation Kit for Cultured Cells (ab110170)
      • Mitochondria Isolation Kit for Cultured Cells (with Dounce Homogenizer) (ab110171)

    Images

    • Western blot - Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer) (ab110169)
      Western blot - Mitochondria Isolation Kit for Tissue (with Dounce Homogenizer) (ab110169)

      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

    • Protocol Booklet

    Click here to view the general protocols

    Datasheets and documents

    • SDS download

    • Datasheet download

      Download

    References (18)

    Publishing research using ab110169? Please let us know so that we can cite the reference in this datasheet.

    ab110169 has been referenced in 18 publications.

    • Liu P  et al. Ferroptosis contributes to isoflurane-induced neurotoxicity and learning and memory impairment. Cell Death Discov 7:72 (2021). PubMed: 33828088
    • Liu Y  et al. Attenuation of Ischemic Stroke-Caused Brain Injury by a Monoamine Oxidase Inhibitor Involves Improved Proteostasis and Reduced Neuroinflammation. Mol Neurobiol 57:937-948 (2020). PubMed: 31620993
    • Gao P  et al. Inhibition of Mitochondrial Calcium Overload by SIRT3 Prevents Obesity- or Age-Related Whitening of Brown Adipose Tissue. Diabetes 69:165-180 (2020). PubMed: 31712319
    • Kawaguchi S  et al. ß3-Adrenergic receptor blockade reduces mortality in endotoxin-induced heart failure by suppressing induced nitric oxide synthase and saving cardiac metabolism. Am J Physiol Heart Circ Physiol 318:H283-H294 (2020). PubMed: 31834837
    • Huang LS  et al. mtDNA Activates cGAS Signaling and Suppresses the YAP-Mediated Endothelial Cell Proliferation Program to Promote Inflammatory Injury. Immunity 52:475-486.e5 (2020). PubMed: 32164878
    View all Publications for this product

    Customer reviews and Q&As

    Show All Reviews Q&A
    Submit a review Submit a question

    1-10 of 22 Abreviews or Q&A

    Question

    Customer kindly contacted us inquiring about sample preparation and storage.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 17 2012

    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.




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

    Read More

    Abcam Scientific Support

    Answered on Oct 18 2012

    Question

    I am removing brains from a number of rats. I'd like to be able to remove all the brains in one go and store them until I'm ready to process the tissue all at the same time. I'd like to use the tissue, once processed using ab110169, for the following kits 1) ab109721, 2) ab110413 and ab109902.
    So the main question is, can I store the brains by freezing (eg. liquid nitrogen to -80 deg C) and thaw the brains and produce extracts that will produce viable sample for use with all 3 kits noted above.

    Read More

    Abcam community

    Verified customer

    Asked on Aug 12 2014

    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.

    Read More

    Jeremy Kasanov

    Abcam Scientific Support

    Answered on Aug 12 2014

    Question

    I am removing brains from a number of rats. I'd like to be able to remove all the brains in one go and store them until I'm ready to process the tissue all at the same time. I'd like to use the tissue, once processed using ab110169, for the following kits 1) ab109721, 2) ab110413 and ab109902.
    So the main question is, can I store the brains by freezing and then thaw the brains and produce extracts that will produce viable sample for use with all 3 kits noted above. Thanks!

    Read More

    Abcam community

    Verified customer

    Asked on Aug 12 2014

    Answer

    Yes, you can use frozen tissues as sample for all three kits. As a reminder, please follow each individual protocol included with each kit to prepare samples for that particular assay.

    Read More

    Jeremy Kasanov

    Abcam Scientific Support

    Answered on Aug 12 2014

    Question

    Is ab110169 suitable also for use with human tissues?

    Does ABCAM guarantee cover the use of ab110169 for that specific application?

    As the ab110169 booklet says it is for "mitochondria isolations from Rat brain, liver and

    heart tissues", is it possible to get a discount code for test in a new application - mitochondria isolations from human tissue (abtrial)?

    Thanks a lot for your attention.

    Read More

    Abcam community

    Verified customer

    Asked on Oct 30 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 30 2012

    Question

    Hi,

    I purchase your mitochondrial isolation kit (MS851) a while ago and have been using it to isolate mitochondria from various arthropods, ie taking a leg an running it through the kit. It is working well and I’m not getting carry over of nuclear DNA. However, when I use rolling circular amplification to replicate the mitochondrial genome I’m getting bacteria contamination. Is there a way to enrich the mitochondria so that the bacteria is removed?

    Cheers

    Read More

    Abcam community

    Verified customer

    Asked on Oct 05 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Oct 05 2012

    Question

    Hola buenos dias,

    Me pongo en contacto con ustedes para comunicarles que en el producto (ab110169) adquirido en Julio de este mismo año venia uno de los dos émbolos (A) roto. El producto no se ha abierto hasta el dia de hoy por lo tanto hasta hoy no hemos tenido noticia del defecto.
    Atentamente,

    Read More

    Abcam community

    Verified customer

    Asked on Sep 07 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Sep 07 2012

    Question

    Hello,

    Just a quick question to see if you do any cell fractionation kits for samples from whole tissue?

    I've noticed ab109718 and ab109719 but they only mention cultured cells?

    Many thanks,

    Read More

    Abcam community

    Verified customer

    Asked on Aug 17 2012

    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/Mitochondria-Isolation-Kit-for-Tissue-ab110168.html (or use the following: https://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-ab110168.html).

    https://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-with-Dounce-Homogenizer-ab110169.html (or use the following: https://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-with-Dounce-Homogenizer-ab110169.html).

    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.

    Read More

    Abcam Scientific Support

    Answered on Aug 17 2012

    Question

    We recently purchased Mitochondria/CytosolFractionation Kit(ab65320) from Abcam. we do not haveDounce tissue grinder, Can we use sonicator toHomogenize cells after addition of1X Cytosol Extraction Buffer Mix.

    please suggest what can i do .

    Read More

    Abcam community

    Verified customer

    Asked on Jul 19 2012

    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/Mitochondria-Isolation-Kit-for-Tissue-with-Dounce-Homogenizer-ab110169.html (or use the following: https://www.abcam.com/Mitochondria-Isolation-Kit-for-Tissue-with-Dounce-Homogenizer-ab110169.html).


    Read More

    Abcam Scientific Support

    Answered on Jul 19 2012

    Question

    Hello dear friends,

    One customer wants to isolate Mitochondria from cardiac tissue, employing your kit ab110169 (MS851).



    The situation is that, they need to check if the protocol that they employ for to treat the samples before the mitochondria isolation could be compatible with your product.



    They treat the tissue for to produce ischemia, and after, they try to do a reoxygenation of it.



    - Ischemia: Krebs Henseillet buffer, pH=6.8 + deoxyglucose + CaCl2

    - Reoxygenation (during 120´): Krebs Henseillet buffer, pH 7.4 + gluose +CaCl2



    The composition of the Krebs Henseillet buffer:







    NaCl



    118mM





    HEPES



    20mM





    MgSO4



    15%







    KH2PO4



    1 M





    KCl



    2 M







    Please, I will be very grateful to you, if you confirm me if your kit could be valid for cardiac tissue, the customer has between 200-400mg of each sample, and other question is, if the Isolation Buffer that you include could be compatible with the Krebs Henseillet buffer.



    Perhaps, do you recommend any specific regarding the previously processing before to do the isolation?, the cardiac tissue is a very connective one.



    Thanks in advance for your collaboration.



    Best Regards,

    Read More

    Abcam community

    Verified customer

    Asked on Jul 16 2012

    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.

    Read More

    Abcam Scientific Support

    Answered on Jul 16 2012

    Question

    The Mitochondria Isolation Kit I would like to request is the one for tissue. I take it there is no difference in SDS between with and without the Dounce Homogenizer?

    Read More

    Abcam community

    Verified customer

    Asked on Jun 06 2012

    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

    Read More

    Abcam Scientific Support

    Answered on Jun 06 2012

    1-10 of 22 Abreviews or Q&A

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