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Thermo Scientific™ Subcellular Protein Fractionation Kit for Cultured Cells
Description
Perform segregation and enrichment of proteins from five different cellular compartments with Thermo Scientific™ Subcellular Protein Fractionation Kit for Cultured Cells.
Highlights:
Concise – extract functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions in less than three hours
Convenient – perform a simple procedure without using ultracentrifugation over gradients
Complete – obtain cytoplasmic, membrane, soluble nuclear, chromatin-bound and cytoskeletal protein fractions from a single kit
Cultured Cells – developed and optimized for use with cultured mammalian cells; for tissue samples use the Subcellular Fractionation Kit for Tissues, Part No. 87790
Compatible – use with a variety of downstream applications including Western blotting, protein assays, electrophoretic mobility shift assays and reporter-gene and enzyme-activity assays
Small-scale – kit provides sufficient reagents to fractionate 50 cell pellets of 20μL each, equivalent to a total of 2g of cell paste or tissue
Includes:
Four extraction buffers, micrococcal nuclease enzyme, calcium chloride solution and protease inhibitor cocktail
Recommended for:
Extraction and enrichment of proteins from different cellular compartments; Separation and profiling of protein from the cytoplasm, nucleus, membrane and cytoskeleton from the same cell sample; Studying protein translocation
Specifications
Specifications
| Product Type | Subcellular Protein Fractionation Kit |
| Content And Storage | Upon receipt store kit at -20°C, or store individual components as indicated above. Kit is shipped with dry ice. |
| Form | Liquid |
| Buffer | Lysis Buffer |
| Reagent Type | Cell Lysis Buffer, Detergent Solution, Subcellular Fractionation |
| Sufficient For | 50 Samples |
| Quantity | 1 Kit |
Frequently Asked Questions (FAQs)
You can choose from several types of Dynabeads magnetic beads for coating with antibody against the subcellular component of choice. We currently do not offer any ready-to-use Dynabeads magnetic beads against a particular subcellular component, and recommend that you choose the beads that work best with your antibody and your application.
The following groups of Dynabeads magnetic beads can be used for this purpose:
(1) Surface Activated Dynabeads magnetic beads
Antibodies from any origin can be conjugated directly to these uncoated beads through covalent bonds. For better orientation of primary antibody on the beads, a secondary antibody (linker antibody) can be coated on the beads first. The Surface Activated Dynabeads magnetic beads are provided in different sizes (4.5, 2.8 and 1 micron) and the larger ones are mostly preferred for this application.
(2) Secondary Coated Dynabeads magnetic beads
These Dynabeads magnetic beads are pre-coated with a secondary antibody and depending on their specificity, they can be easily coated with your primary antibody within a short time period. These beads are provided in two sizes, 4.5 and 2.8 micron. In addition to these beads, our Dynabeads magnetic beads Protein A or Protein G can also be easily coated with antibody from several species. Please see the following links for information about all available Secondary coated Dynabeads magnetic beads and Dynabeads magnetic beads Protein A and Protein G.
(3) Streptavidin Coated Dynabeads magnetic beads
These Dynabeads magnetic beads are coated with streptavidin and require a biotinylated antibody. These beads are provided in two different sizes, 2.8 and 1 micron.
Find more information about Dynabeads magnetic beads at http://www.thermofisher.com/Dynabeads magnetic beads.
The Subcellular Protein Fractionation Kit for Cultured Cells is covered under our general 1-year warranty and is guaranteed to be fully functional for 12 months from the date of shipment, if stored as recommended. Please see section 8.1 of our Terms & Conditions of Sale (https://www.thermofisher.com/content/dam/LifeTech/Documents/PDFs/Terms-and-Conditions-of-Sale.pdf) for more details.
Yes, the nuclear fraction is compatible with co-immunoprecipitation (co-IP) after dialysis, desalting, or 50:50 dilution. The cytoplasmic and membrane fractions are compatible with co-IP directly after extraction and the pellet fraction is compatible with co-IP after detergent removal.
M-PER Mammalian Protein Extraction Reagent can extract some membrane or cytoskeletal proteins, but the extraction efficiency is not consistent. The reagent was not intended to specifically extract these proteins. We recommend using Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) for membrane protein extraction or Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for compartmental extraction.
We offer the following kits for cell fractionation:
Mem-PER Plus Membrane Protein Extraction Kit, Cat. No. 89842
NE-PER Nuclear and Cytoplasmic Extraction Reagents, Cat. No. 78835
Subcellular Protein Fractionation Kit for Cultured Cells, Cat. No. 78840
Subcellular Protein Fractionation Kit for Tissues, Cat. No. 87790
Mitochondrial Isolation Kit for Cultured Cells, Cat. No. 89874
Mitochondrial Isolation kit for Tissue, Cat. No. 89801
Lysosome Enrichment Kit for Tissues and Cultured Cells, Cat. No. 89839
For Research Use Only. Not for use in diagnostic procedures.