text.skipToContent text.skipToNavigation

Qproteome Cell Compartment Kit

For fractionation of proteins according to cellular location
Cat No./ID: 37502
Qproteome Cell Compartment Kit
€280.00
Add To Cart
For up to 10 subcellular fractionations*: Extraction buffers, Protease Inhibitor Solution, Benzonase®.
The Qproteome Cell Compartment Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.
0
Localization of proteins in cells under different growth conditions.
Micrographs (upper panels) of HeLa cells [A] under normal growth conditions and [B] after 4 hour incubation with staurosporine, an inducer of apoptosis. Lower panels show western blots prepared after cell lysates were processed using the Qproteome Cell Compartment Kit. The blots were probed with anti-cytochrome c antibody. It is known that during apoptosis cytochrome c is translocated from the intermembrane space of mitochondria to the cytosol, a process that is reflected in the detection of cytochrome c in the cytosolic fraction of apoptotic cells.
1
Flowchart.
2
Specific separation of marker proteins.
Western blots of fractionated NIH 3T3 cells. Protein (20 µg) from the cytosolic (C), membrane (M), and nuclear (N) fractions was separated by SDS-PAGE. After Western blotting, proteins specific to each fraction were detected using  [A] annexin,  [B] TIM23, and  [C] lamin antibodies, and an HRP-conjugated secondary antibody with chemiluminescent detection.
3
Localization of proteins in tissue samples.
The indicated rat tissue was processed using the Qproteome Cell Compartment Kit. The respective fractions (C: cytosol, M: membrane, N: nucleus, and S: cytoskeleton) were separated by SDS-PAGE and transferred by western blotting to nitrocellulose membrane. The blot was probed with antibodies against the indicated cell-compartment-specific marker proteins.
Performance
Use of the Qproteome Cell Compartment Kit allows simple and effective cell compartment-specific localization of marker proteins from isolated cells (see figure "Specific Separation of Marker Proteins" and to  "Localization of Proteins in Cells Under Different Growth Conditions") or from tissue samples (see figure "Localization of Proteins in Tissue Samples").
Principle

The kit contains 4 Extraction Buffers which enable the sequential isolation of proteins associated with the cytosol, membranes, nucleus, and cytoskeleton from cell lysates or tissues.

Procedure

The 4 Extraction Buffers are added sequentially to a cell pellet and the respective fractions are isolated by centrifugation (see flowchart).

In an alternative protocol for tissues, samples are simultaneously homogenized and disrupted using the TissueRuptor before being processed as for the cell pellet.

Applications

Subcellular fractionation of proteins can be used for the enrichment of low-abundance species; to define the subcellular localization of enzymes, regulatory, and structural proteins; and for monitoring of compartmental redistribution of biomolecules under basal and stimulated conditions.

Features
Specifications
Applications SDS-PAGE, mass spectrometry
Binding capacity/yield Varies
Fractions isolated Four fractions
Sample size ~5 x 10e6 cells
Species Eukaryotes
Start material Cell lysate

You are not authorized to download the resource

fragment fix placeholder

Customers who bought these products also bought