• Product name
    Cell Fractionation Kit - Standard
    See all Cell Fractionation kits
  • Assay type
  • 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 from mammals and other species.

    The Cell Fractionation Kit is designed to produce highly enriched fractions that allow for the monitoring of proteins of interest through various cellular compartments. It is not designed to produce purified cellular fractions.

    This kit does not require mechanical disruption of the sample. This is important as mechanical disruption can disrupt mitochondrial membranes, causing the biologically-irrelevant release of proteins. This kit is 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




  • Samples were loaded as follows from left to right: (1) Marker, (2) whole cell lysate, (3) cytosolic fraction lysate, (4) membrane fraction lysate and (5) nuclear fraction lysate. Membrane was blotted with a Plasma Membrane Fractionation WB cocktail ab140365 containing anti-Sodium Potassium ATPase antibody (110 kDa), anti-GAPDH antibody (37 kDa) and anti-Histone H3 (di methyl K9) antibody.

  • 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.



This product has been referenced in:
  • Paredes F  et al. Poldip2 is an oxygen-sensitive protein that controls PDH and aKGDH lipoylation and activation to support metabolic adaptation in hypoxia and cancer. Proc Natl Acad Sci U S A 115:1789-1794 (2018). Read more (PubMed: 29434038) »
  • Chattaragada MS  et al. FAM49B, a novel regulator of mitochondrial function and integrity that suppresses tumor metastasis. Oncogene 37:697-709 (2018). Read more (PubMed: 29059164) »

See all 30 Publications for this product

Customer reviews and Q&As

Excellent for nuclear extraction

Excellent Excellent 5/5 (Ease of Use)
The kit was used to extract nuclear fraction from whole cell lysates. The mitochondrial fraction was not used. Lamin B1 was used as a nuclear marker and a strong signal was observed in the whole cell extract (WCE) and nuclear fractions (NUC) but not the cytoplasmic fraction (CYTO). Nuclear DNA was only sheared by vortexing and therefore the DNA present in the nuclear sample did cause smearing of the bands. Sonication could improve this but this was enough to demonstrate that the kit could quickly and efficiently purify nuclear fractions.

Abcam user community

Verified customer

Submitted Mar 31 2017

Got clean nuclear fraction

Good Excellent 5/5 (Ease of Use)
The kit is quite easy to use. I got a clean nuclear fraction (N) with positive CREB staining and negative tubulin staining. However, the cytosolic fraction (C) has nuclear contamination (CREB positive).

Ms. Simin Li

Verified customer

Submitted Dec 17 2016

Cell fractionation with Hela cells

Excellent Good 4/5 (Ease of Use)
Follow the protocol in general, but wash with Buffer 1 between different extractions.

Abcam user community

Verified customer

Submitted Jan 14 2016

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: https://www.abcam.com/index.html?datasheet=109719).

The procedure uses only detergen...

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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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