Protein Fractionation and Enrichment Kit (ProFEK, K-0015)
$111.15
ITSIPrep ProFEK (K-0015) provides a validated, fast, and cost-effective method to fractionate and enrich for membrane, cytosol, and nuclear proteins without ultra centrifugation.
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) 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
(20 Assays)
Additional information
| Weight | 2 lbs |
|---|---|
| Dimensions | 12 × 9 × 2 in |
Data Sheet(s)
Click on the title(s) below to view the product data sheet.
Citations
Beta-Catenin Phosphorylated at Threonine 120 Antagonizes Generation of Active Beta-Catenin by Spatial Localization in trans-Golgi Network.
Cheng Du1, Chuanyou Zhang1, Zhuo Li1, Md. Helal Uddin Biswas1, K. C. Balaji2.PLos ONE. 2012 April 12; Volume 7, Issue 4
Article Snippet: “To prepare whole cell lysates, cells cultured on monolayer were washed by phosphoate buffered saline (PBS) and dissolved in RIPA buffer [25] (50 mM Tris-HCl, pH7.4, 150 mM NaCl, 0.5% sodium deoxycholate, 0.1% SDS and 1% NP-40) plus protease inhibitor cocktail (Sigma Chemicals, cat# P-8340). For fractionation experiments, a commercial cell fractionation kit (the ProFEK kit from ITSI Biosciences) was used according to manufacturer’s recommendation.“
Sample Preparation for 2D Electrophoresis and CE-Based Proteomics.
J. M. Nagy, A. Lipka, F. Pereira, N. Marlin, S. Hassard Springer Science+Buisness Media (google book) 2011.
Article Snippet: “Sample solubilisation can be achieved using these conventional reagents (see Table 3.1) or commercially available kits, (ZOOM® 2D Protein Solubilizer Kit (Invitrogen); Bio-X Hydrophobic Protein Solubilizer for 2D Electrophoresis (InfiniteBio Inc.); FOCUST™ Protein Reduction-Alkylation Kit (Biosciences), ProteoPrep® Reduction & Alkylation Kit (Sigma-Aldrich) and Protein Fraction Enrichment Kit (ProFEK) from ITSI Biosciences) with specific techniques applicable to a range of protein analyses.“
Characterization of membranous and cytoplasmic EGFR expression in human normal renal cortex and renal cell carcinoma.
Yeong-Shiau Pu, Chao-Yuan Huang, Yi-Zih Kuo, Wang-Yi Kang, Guang- Yaw Liu, A-Mei Huang, Hong-Jeng Yu, Ming-Kuen Lai1, Shu-Pin Huang, Wen-Jeng Wu, Shean-Jaw Chiou, Tzyh-Chyuan Hou Int. J Oncol. 2009 September;16:82.
Article Snippet: “The cytoplasmic and membranous proteins were extracted from tissue samples using the ITSIPREP™ ProFEK KIT (ITIS Bioscience, PA, USA). About 50-100 mg of tissue was used and cut into small pieces.“
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