Biological samples contain a complex mixture of proteins, which in living cells are localized in the membrane, nucleus and/or cytoplasm. Therefore, to identify the proteins that are predominant in each region, it is necessary to reduce the complexity of the total protein lysate by fractionating the lysate into membrane, nuclear and cytosol fractions. Reduction in sample complexity is also important when the detection of protein(s) that occur at low concentrations in any of the regions is desired. An ideal method should be reproducible, simple, convenient and cost effective.
ITSI has a unique kit for fractionation and enrichment of proteins found in the nuclear, cytosol and membrane regions. The ITSIPrep ProFEK (K-0015) Protein Fraction and Enrichment Kit (ProFEK) provides a validated, fast, and cost-effective method to fractionate and enrich for membrane, cytosol, and nuclear proteins without ultra centrifugation. The optimized reagents and procedure provided with the kit allow for reproducible results and identification of unique region associated proteins. The extracted membrane, nuclear, and/or cytoplasmic protein fractions can be analyzed by mass spectrometry, SDS-PAGE (Fig 1), 2D-PAGE, 2D-DIGE (Fig 2), and western blotting.
Fig 1: 1D-PAGE gel image of chicken heart protein lysate fractionated with ITSIPrep ProFEK kit into Nuclear (lane 1), Cytosol (lane 2) and Membrane (lane 3) protein fractions. A and B represent protein fractions obtained with 1x and 2x volume of solvent respectively, MWW is molecular weight marker and WT is Whole (unfractionated) tissue lysate.
Benefits of using the ITSIPrep Protein Fractionation and Enrichment Kits:
– Lowest cost/sample for selective enrichment of nuclear, cytosol and membrane proteins
– No ultra centrifugation required
– Reproducible results
– Easy-to-follow protocol
Fig 2: Images showing proteins from human tissue lysate fractionated with ITSIPrep ProFEK kit to concentrate Nuclear, Cytosol and Membrane proteins prior to analysis by 2D-DIGE
Each Kit contains optimized reagents and consumables including:
- ProFEK Buffer 1
- ProFEK Buffer 2
- ProFEK Buffer 3
- Disposable Pestle/Tube for sample homogenization
- Standard protocol
K-0015-20 (20 Assays) – $97.50