539790
ProteoExtract® Subcellular Proteome Extraction Kit
Szinonimák:
S-PEK Kit
Bejelentkezésa Szervezeti és Szerződéses árazás megtekintéséhez
Összes fotó(1)
About This Item
UNSPSC kód:
41106500
NACRES:
NA.77
Javasolt termékek
használat
sufficient for 20 extractions
gyártó/kereskedő neve
Calbiochem®
tárolási körülmény
OK to freeze
avoid repeated freeze/thaw cycles
technika/technikák
fractionation: suitable
bemenet
sample type: mammalian tissue(s)
sample type: mammalian cultured cells
kiszállítva
ambient
tárolási hőmérséklet
2-8°C
Általános leírás
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:
Sample size: 3-5x106 or 25-50 mg tissue.
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)
Sample size: 3-5x106 or 25-50 mg tissue.
Tulajdonságok és előnyök
- 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
Komponensek
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.
Figyelmeztetés
Toxicity: Multiple Toxicity Values, refer to MSDS (O)
Elv
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.
Elkészítési megjegyzés
Kit Components Needed for One Extraction
The volume of each component required for one subcellular extraction depends on the amount of starting material.
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
Tárolás és stabilitás
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.
Egyéb megjegyzések
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
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
Jogi információk
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
Veszélyességi osztályok
Aquatic Chronic 3 - Eye Irrit. 2 - Skin Irrit. 2
WGK
WGK 3
Analitikai tanúsítványok (COA)
Analitikai tanúsítványok (COA) keresése a termék sarzs-/tételszámának megadásával. A sarzs- és tételszámok a termék címkéjén találhatók, a „Lot” vagy „Batch” szavak után.
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