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539790

Millipore

ProteoExtract® Subcellular Proteome Extraction Kit

Synonyme(s) :

S-PEK Kit

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About This Item

Code UNSPSC :
41106500
Nomenclature NACRES :
NA.77

Utilisation

sufficient for 20 extractions

Fabricant/nom de marque

Calbiochem®

Conditions de stockage

OK to freeze
avoid repeated freeze/thaw cycles

Technique(s)

fractionation: suitable

Entrée

sample type: mammalian tissue(s)
sample type: mammalian cultured cells

Conditions d'expédition

ambient

Température de stockage

2-8°C

Catégories apparentées

Description générale

Fast and reproducible extraction of subcellular proteomes from mammalian cells

ProteoExtract® Subcellular Proteome Extraction Kit (S-PEK) is designed for fast and reproducible extraction of subcellular proteomes from adherent and suspension-grown mammalian cells. The S-PEK takes advantage of the different solubilities of certain subcellular compartments in the four selected reagents. In the case of adherent cells, the procedure is performed directly in the tissue culture dish without the need for cell removal. Cells or the parts of the cells remain attached to the plate during sequential extraction of subcellular compartments, until the appropriate extraction reagent is used. Thus, the early destruction of the cellular structure by enzymatic or mechanical detachment of cells from the tissue culture plate and any mixing of different subcellular compartments is prevented. For suspension-grown cells, extraction starts with gentle sedimentation and washing of the cells. The stepwise extraction delivers four distinct protein fractions from one sample:


  • Cytosolic fraction (F1)
  • Membrane/organelle protein fraction (F2)
  • Nucleic protein fraction (F3)
  • Cytoskeletal fraction (F4)
Proteins are obtained in the native state making the S-PEK suitable for many downstream applications such as 1D and 2D gel electrophoresis, immunoblotting, enzyme activity assays, and protein microarrays.
Sample size: 3-5x106 or 25-50 mg tissue.

Caractéristiques et avantages

  • Stepwise extraction resulting in four distinct subcellular proteomes from one sample
  • Highly reproducible
  • No ultracentrifugation steps
  • Fast—needs just 2 hours with 45 minutes hands-on time
  • Produces proteins suitable for functional studies

Composants

Wash buffer, Extraction Buffer I, Extraction Buffer II, Extraction Buffer III, Extraction Buffer IV, Protease Inhibitor Cocktail, Benzonase® Nuclease (Cat. No. 71206), and a user protocol.

Avertissement

Toxicity: Multiple Toxicity Values, refer to MSDS (O)

Principe

The Calbiochem ProteoExtract Subcellular Proteome Extraction Kit is designed for the subcellular extraction of mammalian proteins from the cytosolic, organelle and membrane, nuclear, and cytoskeletal fractions of adherent tissue culture cells, suspension tissue culture cells, frozen cell pellets, and fragmented tissues.

Notes préparatoires

Kit Components Needed for One Extraction


The volume of each component required for one subcellular extraction depends on the amount of starting material.
Appropriate samples types include:
  • Adherent tissue culture cells
  • Suspension-grown tissue culture cells
  • Frozen cell pellets
  • Fragmented tissue

Stockage et stabilité

Prior to performing the extraction protocol all frozen buffers must thawed at room temperature. A water bath set at 25°C will aid in the thawing process. After thawing, mix the buffers well gentle shaking or vortexing.

Autres remarques

Zhang, L., and Insel, P. A. 2004. J. Biol. Chem.279, 20858.
Yuan, X., et al. 2002. Electrophoresis23, 1185.
Butcher, et al. 2001. J. Immunol.167, 2193.
Ott, et al. 2001. Pharmacogenomics J.1, 142.
Allen, L. 2000. Nature405, 819.
Dunn, M. J. 2000. Electrophoresis 6.
Rabilloud, T. 2000. Two-dimensional Gel Electrophoresis and Identification Methods Springer-Verlag
Mejdoubi, et al. 1999. Biochem. Biophys. Res. Comm.254, 93.
Reymond, et al. 1997. Electrophoresis18, 2842.
Laemmli, U. K. 1970. Nature227, 680.
Lowry, et al. 1951. J. Biol. Chem.193, 265.

http://www.expasy.ch/ and http://www.expasy.proteome.org.au

Informations légales

Benzonase is a registered trademark of Merck KGaA, Darmstadt, Germany
CALBIOCHEM is a registered trademark of Merck KGaA, Darmstadt, Germany
PROTEOEXTRACT is a registered trademark of Merck KGaA, Darmstadt, Germany

Classification des risques

Aquatic Chronic 3 - Eye Irrit. 2 - Skin Irrit. 2

Classe de danger pour l'eau (WGK)

WGK 3

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