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Subcellular Protein Extraction
One of the
challenges of functional proteomics is separation of
complex protein mixtures for quantitative and
differential subcellular localization analysis. This
necessitates standardized and repeatable operation
procedures to isolate subcellular proteomes from
tissues and cells. BioChain¡¯s CNMCS and CNM
Compartmental Protein Extraction Kits address the
challenge by providing an innovative,
easy-to-perform, and cost-effective method to
sequentially isolate cytoplasmic, nuclear, membrane,
and cytoskeleton proteins from mammalian tissues and
cells based on a proprietary technique. These unique
kits can isolate four or three compartmental
proteins. They are excellent tools for the initial
purification and preparation of proteins for
downstream applications including SDS-PAGE, Western
blotting, gel mobility shift, protein assays and
other procedures.
Features
¡ñ Convenient -
Provides a complete set of components for stepwise
preparation of cytoplasmic, nuclear, and membrane
proteins from mammalian tissues and cultured cells
¡ñ Fast - isolate four or three compartmental
proteins in less than three hours
¡ñ Reliable - Super-quality and highly reproducible
separation of subcellular proteome fractions as
verified by clearly distinct protein patterns and
precise subcellular localizations of marker proteins
¡ñ Pure - Minimal cross contaminations
¡ñ Simple - Easy
to use comparing to other methods, such as
differential centrifugations
Application
The Subcellular Protein Extraction Kit can be used
for:
¡ñ Detection of differential post-translational
modifications or differential subcellular
localization of target proteins
¡ñ Enrichment of low-abundance proteins for
visualization and subsequent analysis
¡ñ Preparation of nuclear extract from mammalian
tissues and cultured cells is crucial for studying
DNA binding proteins such as transcription factors
employing gel mobility shift techniques
¡ñ Preparations
of cytoplasmic fractions are useful to study soluble
proteins abundant in cytosol
¡ñ
Preparations of
membrane fractions to study membrane proteins such
as receptors
Quality
Control
One kit of this lot has been tested to go through
the complete compartment protein extraction
procedure from rat colon. Four compartmental
proteins from the extraction are used for
electrophoresis, transferring to PVDF membrane and
immunoblotting with Mouse anti-GAPDH, Mouse anti-Histone
H1, Mouse anti-Na+/K+ ATPase, and Mouse anti-Vimentin
as primary antibodies, and HRP-conjugated anti-Mouse
IgG as secondary antibody. The separation of four
compartmental proteins is confirmed by the
enrichment of GAPDH in the cytoplasmic fraction,
Histone H1 in the nuclear fraction, Na+/K+ ATPase in
the membrane fraction, and the unique localization
of Vimentin in the cytoskeleton fraction |
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