蛋白组分抽提试剂盒说明书

Overview

∙Description

∙References

∙Product Information

∙Applications

∙Biological Information

∙Safety Information

∙Product Usage Statements

∙Storage and Shipping Information

∙Supplemental Information

∙Prix & Disponibilité

Prix & Disponibilité

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Overview

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 subce

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

the cellular structure by enzymatic or mechanical detachment of cells from the tissue cultu

plate and any mixing of different subcellular compartments is prevented. For

suspension-grown cells, extraction starts with gentle sedimentation and washing of the cel

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 assa

and protein microarrays.

Sample size: 3-5x106or 25-50 mg tissue.

Catalogue

Number

539790

Brand

Family

Calbiochem®

Synonyms S-PEK Kit

Features and benefits ∙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

Application

Data

A:Adherent SAOS cells were extracted according to the Detailed Protocol

Subcellular Extraction of Proteins From Adherent Cells as outlined above. Im

i-iv show the morphology of the cells before and after each extraction s

(200-fold enlarged). The SAOS cells remained attached throughout the extrac

procedure. B:An aliquot of each fraction from A were subjected to SDS-

analysis (F1-F4 = fractions 1-4). The data demonstrate clear differences in

protein banding patterns among the 4 fractions. C:Aliquots of each frac

from A were separated by SDS-PAGE and transferred to PVD membrane for blot

with the indicated antibodies. For c-Fos, the fractions were subjected t

immunoprecipitation, prior to Western blotting, to enrich for any c-Fos pre

in each fraction. The data demonstrate that each marker protein is specific

enriched within the appropriate fraction.