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Thermo Scientific™ Subcellular Protein Fractionation Kit for Cultured Cells

Catalog No. PI78840
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Catalog No. PI78840 Supplier Thermo Scientific™ Supplier No. 78840
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Description

Enrich for cultured cell-specific cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal proteins in less than 3 hours.

Perform segregation and enrichment of proteins from five different cellular compartments with Thermo Scientific™ Subcellular Protein Fractionation Kit for Cultured Cells.

The kit enables the stepwise separation and extraction of cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal proteins from mammalian cultured cells. Extracts obtained with the Subcellular Protein Fractionation Kit for Cultured Cells are compatible with a variety of downstream applications including Western blotting, protein assays, electrophoretic mobility shift assays and reporter-gene and enzyme-activity assays. Fractionation of subcellular proteins enables protein localization assessment and protein enrichment from specific cellular compartments. The Thermo Scientific Subcellular Protein Fractionation Kit includes a combination of reagents for stepwise lysis of cells into functional cytoplasmic, membrane, nuclear soluble, chromatin-bound and cytoskeletal protein fractions in less than 3 hours. Extracts from each subcellular compartment generally have less than 15% contamination between fractions, which is sufficient purity for most experiments studying protein localization and redistribution.

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

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)

What products are available for the isolation of subcellular components?

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.
Can you provide the shelf-life for the Subcellular Protein Fractionation Kit for Cultured Cells?

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.
Can I use the Subcellular Protein Fractionation Kit for Cultured Cells (Cat. No. 78840) for subsequent co-immunoprecipitation (co-IP) analysis of different nuclear proteins?

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.
Will M-PER Mammalian Protein Extraction Reagent extract membrane or cytoskeletal proteins?

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.

What kits do you offer for cell fractionation?

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
Why is cell fractionation important?

Isolation of subcellular fractions and concentration of low-abundantce proteins allows for more efficient identification and study of the proteins of interest.

For Research Use Only. Not for use in diagnostic procedures.