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Subcellular Protein Fractionation Kit

Catalog # NBP2-47659 | Novus Biologicals a Bio-Techne Brand
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NBP2-47659

Key Product Details

Species

Mammal

Applications

Enzyme Activity, Gel Shift, Western Blot

Kit Type

Fractionation Kit

Product Summary for Subcellular Protein Fractionation Kit

The Cell Fractionation kit is designed to provide reproducible extraction of four subcellular protein fractions (cytosol, nucleus, membrane/particulate, and cytoskeletal fractions) from a single mammalian sample. The method is fast and simple, needing only 2 hours and no ultracentrifugation involved. All four protein fractions obtained are suitable for many downstream applications such as 1-D or 2-D gel, enzyme activity assays, gel shift assay, and Western blotting.

The percentage of proteins in the different fractions are cell-dependent, usually in the range of 20-30% cytosol, 25-35% membrane (organelles), 15-25% Nuclei, 15-25% insoluble.

This product is manufactured by Abcam and distributed by Novus Biologicals. (Abcam Catalog Number: ab288085)

Product Specifications

Application Notes

In Western analysis, if a membrane protein appeares mainly in the nuclear fraction, the membrane extraction step may not be sufficient. Make sure to add Membrane Extraction Buffer B into the extraction, which contains NP-40 detergent. If it still shows the same result, increase incubation time from 1 min to 5 min after adding MEB-B. NP-40 should extract membrane proteins from most of cells.

Scientific Data Images for Subcellular Protein Fractionation Kit

Subcellular Protein Fractionation Kit [NBP2-47659]

Kit Contents for Subcellular Protein Fractionation Kit

  • Cytosol Extraction Buffer (CEB)
  • DTT (1 M)
  • Membrane Extraction Buffer-A (MEB-A)
  • Membrane Extraction Buffer-B (MEB-B)
  • Nuclear Extraction Buffer (NEB)
  • Protease Inhibitor Cocktail (lyophilized)

Formulation, Preparation, and Storage

Concentration

Concentration is not relevant for this product. Please see the protocols for proper use of this product.

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Storage

Store at -20C. Avoid freeze-thaw cycles.

Product Documents for Subcellular Protein Fractionation Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Product Specific Notices for Subcellular Protein Fractionation Kit

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Kits are guaranteed for 6 months from date of receipt.

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  • Subcellular Protein Fractionation Kit
    Name: Babak Nami
    Verified Customer | Posted 10/23/2018
    This kit works very well mammalian cell lines. The procedure is quite simple and fast. Results in very clean membrane, cytosol and nuclear fractionation but cytoskeletal fraction is contaminated by nuclear fraction. The only problem is that it seems that all fractions are contaminated by cytosol. I got a lot of a-tubulin in the all fractions!!! Very strong a-tubulin band in membrane fraction. a-tubulin is supposed to be cytosolic protein. Having it in cytoskeletal fraction makes sense but not in membrane and nuclear fraction. In my manual protocol I used to use I get tubulin-free membrane and nuclear fractions.
    Subcellular Protein Fractionation Kit NBP2-47659

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