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539790

Millipore

ProteoExtract® Subcellular Proteome Extraction Kit

Sinonimo/i:

S-PEK Kit

Autenticatiper visualizzare i prezzi riservati alla tua organizzazione & contrattuali


About This Item

Codice UNSPSC:
41106500
NACRES:
NA.77

impiego

sufficient for 20 extractions

Produttore/marchio commerciale

Calbiochem®

Condizioni di stoccaggio

OK to freeze
avoid repeated freeze/thaw cycles

tecniche

fractionation: suitable

input

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

Condizioni di spedizione

ambient

Temperatura di conservazione

2-8°C

Descrizione generale

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.

Caratteristiche e vantaggi

  • 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

Componenti

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.

Attenzione

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

Principio

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.

Nota sulla preparazione

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

Stoccaggio e stabilità

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.

Altre note

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

Note legali

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

Classi di pericolo

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

Classe di pericolosità dell'acqua (WGK)

WGK 3


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