Overview

  • Product nameCell Fractionation Kit - Standard
    See all Cell Fractionation kits
  • Assay typeDirect
  • Species reactivity
    Reacts with: Human, a wide range of other species
  • Product overview

    Abcam's Cell Fractionation Kit (Standard) allows for the rapid and simple preparation of mitochondrial, cytoplasmic and nuclear containing fractions from cultured cells. Unlike competing products, this kit does not require mechanical disruption of the sample. This is an important performance characteristic, as mechanical disruption can also disrupt mitochondrial membranes, causing the biologically-irrelevant release of proteins. Particularly useful in studying apoptosis and the movement of pro-apoptotic proteins such as cytochrome c.

     

    Sufficient materials are provided for fractionation of 1 x108 cells or for preparation of 40 samples, each corresponding to one 100 mm plate at 2.5 x 106 cells/plate.

     

    A companion product is also available which allows for the fractionation of cells grown in 96-well plates: Cell Fractionation Kit HT (ab109718).

     

     

  • Notes

    The Cell Fractionation Kit is not designed to produce purified cellualr fractions, there will always be a degree of contanmination between the 3 fractions.

Properties

  • Alternative names
    • MS861

Cell Fractionation Kit - Standard images

  • Cytosolic (C), mitochondrial (M) and nuclear (N) fractions of HepG2 cells were prepared as described in the Protocol. Fractions were analyzed by Western blotting using Abcam's ApoTrack™ Cytochrome c Apoptosis WB Antibody Cocktail (ab110415) containing antibodies against mitochondrial matrix (pyruvate dehydrogenase subunit E1α, PDH E1α), mitochondrial inner membrane (F1-ATPase α), mitochondrial intermembrane space (cytochrome c) and cytosolic (glyceraldehyde-3-phosphate dehydrogenase, GAPDH) markers as well as with antibodies against additional mitochondrial matrix (Hsp70) and nuclear (poly (ADP-ribose) polymerase, PARP and SP1) markers, followed by appropriate HRP-conjugated goat secondary antibodies and ECL detection.

  • In this experiment, apoptosis was induced in Jurkat, HeLa and 143B osteosarcoma cells by treatment with staurosporine or in Jurkat cells by FAS. Mitochondrial and cytoplasmic fractions were isolated (using ab109719) and probed using ab110415. As is clear from the gels, cytochrome c has translocated partially in FAS-induced cells and STS-treated osteosarcoma cells, and almost completely in STS-treated Jurkat and HeLa cells. The three control targets allow for verification of the "cleanness" of the cell fractionation.

Protocols

References for Cell Fractionation Kit - Standard (ab109719)

This product has been referenced in:
  • Abbas W  et al. Blockade of BFA-mediated apoptosis in macrophages by the HIV-1 Nef protein. Cell Death Dis 5:e1080 (2014). Human . Read more (PubMed: 24556695) »
  • Liu B  et al. Apoptosis and pro-inflammatory cytokine response of mast cells induced by influenza A viruses. PLoS One 9:e100109 (2014). Read more (PubMed: 24923273) »

See all 15 Publications for this product

Product Wall



I an confirm that the ab109719 is a cell fractionation kit and it has the word standard in its name to distinguish this product from ab109718 (Cell Fractionation Kit –HT), where the HT stands for high throughput. I believe that we did n...

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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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The kit we recommend for preparing subcellular fractions is ab109719 (or ab109718 for cells in 96-well plates).

Click here (or use the following: http://www.abcam.com/index.html?datasheet=109719).

The procedure uses only detergent...

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The product is guaranteed to work with the cell concentrations as set in the protocol. However, it is certainly possible to create fractions of higher protein concentrations. This may need however some optimization. I would suggest to customer to try t...

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Thank you for contacting us and your interest in our products. I am sorry for the delay in getting back to you.
I have consulted with the lab over the suitability of the two isolation kits, ab109719 and ab110170, in regards to your customers probl...

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The appropriate fractionation depends on the ratio of detergents to the cellular mass. Since cells vary in their size, the extraction condition may need a optimization for a particular cell line. The customer reports good separation of cytoso...

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This product was updated in September 2012 to change the Buffer A from 2 bottles of 1X, to 1 bottle of 2X to streamline the packaging. I will inquire with product updates to see why the protocol was not changed. The 2X is simply diluted 1:1 with H2O...

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Thank you for your inquiry.

1.) I am happy to confirm that cells can be scraped directly in Buffer A. Alternatively, the cells can be scraped in PBS and washed in Buffer A.

2.) Buffer A, B and C are compatible with BCA protein assay...

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Only the SDS PAGE buffer contains DTT, and this buffer can be replaced easily with an alternative sample buffer if desired.

Thank you for your enquiry.

We do not have set expiration dates for our products. We guarantee all of our products to work for at least six month un-opened from the date of purchase when stored correctly.

Please refer to the protoc...

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