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Thermo Scientific™ Mitochondria Isolation Kit for Cultured Cells

Catalog No. PI89874
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PI89874 115 mL
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Catalog No. PI89874 Supplier Thermo Scientific™ Supplier No. 89874
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Description

Isolate intact mitochondria from cultured mammalian cell samples in about 40 minutes with this versatile, benchtop microcentrifuge-based fractionation kit.

Thermo Scientific™ Mitochondria Isolation Kit for Cultured Cells provides a versatile, microcentrifuge-tube method for fractionating intact mitochondria from cultured mammalian cell samples in about 40 minutes.

Isolation of mitochondria is typically a laborious process requiring single-sample processing with Dounce homogenization. The Mitochondria Isolation Kit uses a non-mechanical, reagent-based method that allows multiple (six) cultured cell samples to be processed concurrently in about 40 minutes. Cultured mammalian cell pellets are gently lysed using a proprietary formulation that results in maximum yield of mitochondria with minimal damage to integrity. The product instructions describe guidelines for optimizing purity vs. yield parameters. The kit also offers an optimized Dounce homogenization procedure, which results in two-fold greater mitochondria recovery compared to the reagent-based method. Both methods use differential centrifugation to separate the mitochondrial and cytosolic fractions with a benchtop microcentrifuge and are completed in approximately 40 minutes (post-cell harvest). Once isolated, the mitochondria can be used in downstream applications such as apoptosis, signal transduction and metabolic studies, as well as to facilitate mitochondrial proteomics efforts.

Highlights:

  • Fast – isolate intact mitochondria from 2 × 107 cells in approximately 40 minutes (post-cell harvest)

  • Flexible – process several samples simultaneously using the reagent-based method, or obtain highest possible yield with the Dounce homogenization method

  • Optimizable – guidelines provided for optimizing purity vs. yield with the reagent-based method

  • Benchtop-easy – isolation performed in a microcentrifuge tube

  • Compatible – isolated mitochondria may be lysed with detergent or processed for almost any downstream application, including 2D electrophoresis

Includes:

Three buffer and detergent solutions totaling approx. 115mL

Specifications

Product Type Cell Isolation Kit
Content And Storage Upon receipt store kit at 4°C. Product is shipped at ambient temperature.
Form Liquid
Reagent Type Organelle Isolation, Subcellular Fractionation
Sufficient For 50 Isolations
Quantity 115 mL

Frequently Asked Questions (FAQs)

Can you provide the shelf-life for the Mitochondria Isolation Kit for Cultured Cells?

The Mitochondria Isolation Kit for Cultured Cells is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.
What is a typical yield from the Mitochondria Isolation Kit for Cultured Cells?

The typical yield is approximately 150 µg protein from 20 million starting cells after mitochondrial isolation and lysis, based on HeLa cells. This varies with cell type.
Can I use a Dounce homogenization procedure in conjunction with the Mitochondria Isolation Kit for Cultured Cells (Cat. No. 89874)?

Yes, the kit offers an optimized Dounce homogenization procedure, which results in two-fold greater mitochondri recovery compared to reagent-based method.
Can I further purify my crude mitochondrial fraction isolated using your Mitochondria Isolation Kit for Cultured Cells (Cat. No. 89874)?

To obtain a more purified fraction of heavy mitochondria, centrifuge the post-nuclear supernatant at 3000g rather than 12,000g. Western blot analysis of purified versus crude mitochondrial fractions prepared with this kit result in a > 50% reduction in cathepsin S (lysosomal protein) and PMP70 (peroxisomal protein) in the purified fraction.

What kits do you offer for cell fractionation?

We offer the following kits for cell fractionation:

Mem-PER Plus Membrane Protein Extraction Kit, Cat. No. 89842
NE-PER Nuclear and Cytoplasmic Extraction Reagents, Cat. No. 78835
Subcellular Protein Fractionation Kit for Cultured Cells, Cat. No. 78840
Subcellular Protein Fractionation Kit for Tissues, Cat. No. 87790
Mitochondrial Isolation Kit for Cultured Cells, Cat. No. 89874
Mitochondrial Isolation kit for Tissue, Cat. No. 89801
Lysosome Enrichment Kit for Tissues and Cultured Cells, Cat. No. 89839
Why is cell fractionation important?

Isolation of subcellular fractions and concentration of low-abundantce proteins allows for more efficient identification and study of the proteins of interest.

For Research Use Only. Not for use in diagnostic procedures.