MitoSciences (MS852)

Mitochondria Isolation Kit for Cultured Cells (ab110170)

Overview

  • Product nameMitochondria Isolation Kit for Cultured Cells
    See all Mitochondria Isolation kits
  • Assay typeQuantitative
  • Species reactivity
    Reacts with: Human, a wide range of other species
  • Product overview

    ab110170 contains sufficient reagents for up to 20 benchtop mitochondria isolations from mammalian cultured cells. Isolated mitochondria can be analyzed by Western Blotting, Immunocapture, or Activity assay.

     

    Available with a dounce homogenizer as kit ab110171.

     

  • Tested applicationsFunctional Studiesmore details

Properties

    Applications

    Our Abpromise guarantee covers the use of ab110170 in the following tested applications.

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

    Application Abreviews Notes
    Functional Studies Use at an assay dependent dilution.

    Mitochondria Isolation Kit for Cultured Cells images

    • Isolated mitochondria show enriched signal when compared to the whole cell extract. In lane 1, MRC5 mitochondria isolated with Abcam's ab110170 (MS852) Mitochondria Isolation Kit for Cultured Cells were loaded at 20 µg. In lanes 2 and 3, post-spin supernatant (SN) and whole cell extract were loaded at 20 µg.

    Protocols

    References for Mitochondria Isolation Kit for Cultured Cells (ab110170)

    This product has been referenced in:
    • Silva DF  et al. Bioenergetic flux, mitochondrial mass and mitochondrial morphology dynamics in AD and MCI cybrid cell lines. Hum Mol Genet 22:3931-46 (2013). Read more (PubMed: 23740939) »
    • Gohil VM  et al. Meclizine inhibits mitochondrial respiration through direct targeting of cytosolic phosphoethanolamine metabolism. J Biol Chem 288:35387-95 (2013). Read more (PubMed: 24142790) »

    See all 8 Publications for this product

    Product Wall

    The mitotox kits will work with mitochondria from any human tissue. Therefore it will work with mitochondria from human PBMCs.


    To isolate mitochondria from human PBMCs I suggest the customer to use ab110170 (Mitochondria Isolation Kit f...

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    You can use the leftover reagents of kit ab110170 with tissues. However, you will need to follow the tissue protocol to get the best outcome.

    Note, we have a specific Mitochondrial isolation kit for tissue: ab110168.

    Large clearance pestle (A)[clearance 0.0030-0.0050 in] is used for the initial sample reduction, such as tissue prep;

    Small clearance pestle (B) [0.0005-0.0025 in] is used to form the final homogenate.

    When prepare cell homogenate...

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    The procedure can accomodate approximately 20 benchtop mitochondria isolations, assuming about 4x10^7 cells per isolation are used. These amounts can be scaled up or down accordingly, as long as the cells are resuspended to 5 mg/ml in Reagent A, a...

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    ab109719 is not suitable for this purpose, this kit is based on detergent fractionation of cellular proteins into three fractions. RNA distribution was not tested with this kit.



    Mitochondrial isolation kits (ab110168 – ab1101...

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    I can confirm that the mitochondria isolated using this kit can be used in other biological assays.
    Please check protocol booklet page number 2 for expected yield based on cell line:
    Sample / Starting Material / Expected Yield
    MRC-5 / 4 – ...

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    You will want to use either ab110170 or ab110171 (which includes the homogenizer), as these are optimized for isolating fractions from cultured cells versus tissue samples. The supernatant from SPIN2 is the cytosolic fraction, though it might be contam...

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    Red blood cells do not contain mitochondria so PBMC isolation is recommended.

    Ab110170 can be used to make all of the fractions you need. After the Spin 2 step in the protocol, the supernatant is the cytosolic fraction. The mitochondrial pellet will be largely intact mitochondria. In order to rupture the mitochondria, you can us...

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    With regards to your questions:
    1. What is a cell lifter? Can cells be detached by regular trypsinization (3 min at 37 degrees)?
    A cell lifter is a tool used for scraping cells from dishes and plates.
    Such as this one available through S...

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