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Thermo Scientific™ Mem-PER™ Plus Membrane Protein Extraction Kit

Catalog No. PI89842
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PI89842 300 mL
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Catalog No. PI89842 Supplier Thermo Scientific™ Supplier No. 89842
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Description

Perform small-scale solubilization and enrichment of integral membrane proteins and membrane-associated proteins in a simple reagent-based procedure.

Perform small-scale solubilization and enrichment of integral membrane proteins and membrane-associated proteins in a simple reagent-based procedure using Thermo Scientific™ Mem-PER Plus Membrane Protein Extraction Kit.

Additional Information and Online Ordering

The Mem-PER Plus Kit effectively isolates membrane proteins from cultured cells and tissues using a simple benchtop microcentrifuge procedure. Integral membrane proteins having one or two membrane-spanning domains are effectively solubilized and isolated from cytosolic proteins. The kit makes possible the extraction and selective enrichment of integral and attached membrane proteins from cultured mammalian cells, as well as from hard and soft mammalian tissues. The Mem-PER Plus Kit has many advantages compared to our original Mem-PER Kit. The protocol is simpler, and the fractions are directly compatible with many downstream applications such as SDS-PAGE, Western Blotting, BCA, immunoprecipitation, and amine-reactive protein labeling techniques.

Highlights:

  • Extraction and isolation – produces minimal cross-contamination (typically less than 10%) of cytosolic protein into the membrane protein fraction

  • Cells or tissues – effective for extraction from cultured mammalian cells and mammalian tissues

  • Downstream compatibility – analyze membrane protein extracts by SDS-PAGE, Western blotting, immunoprecipitation and protein assays

  • Fast and simple – isolation of membrane proteins in approximately one hour

  • No special equipment required – only a benchtop microcentrifuge, tubes, homogenizers, and pipettors are needed

Includes:

Kit contains a cell wash solution, permeabilization buffer, and solubilization buffer.

Specifications

Product Type Protein Extraction Kit
Content And Storage Upon receipt store Cell Wash Solution and Solubilization Buffer at 4°C. Aliquot and store Permeabilization Buffer at -20°C to avoid freeze/thaws. Product is shipped on ice.
Form Liquid
Product Line Mem-PER
Reagent Type Cell Lysis Buffer, Detergent Solution, Subcellular Fractionation
Sufficient For 50 Samples
Quantity 300 mL

Frequently Asked Questions (FAQs)

I am using the Mem-PER Plus Membrane Protein Extraction Kit with less than 5 X 10E6 cells. Should I scale down the amount of reagents accordingly?

The kit reagents can be scaled up or down as long as the ratio of reagents to cells is maintained. However, we don't have specific guidelines for the range of cell numbers that can be used because protein yields can vary depending on the cell line. It is important to note that the process of scaling the kit reagents up or down and determining the appropriate cell numbers for optimal protein yield will require empirical optimization by the investigator.
What kind of detergent does the Permeabilization Buffer from the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No 89842) contain?

The composition of the Permeabilization Buffer, including the identity and concentration of any detergent therein, is proprietary. However, the Permeabilization Buffer contains a very small amount of a weak non-ionic detergent to facilitate the permeabilization of the cells and release of the cytosolic proteins.
Will the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) extract membrane proteins from the plasma membrane only or also from other intra-cellular membranes (e.g., nuclear, mitochondrial, endoplasmic reticulum)?

The Permeabilization buffer in the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) creates pores in the plasma membrane of cells that releases cytosolic proteins. The permeabilized cells are centrifuged, and the supernatant contains the cytosolic protein fraction. The pellet is then resuspended in the Solubilization buffer containing a proprietary detergent that solubilizes plasma, mitochondrial, and ER membrane proteins. Subsequent centrifugation yields a supernatant containing solubilized membrane and membrane-associated proteins.
Can microalgae be processed with the Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842)?

The Mem-PER Plus Membrane Protein Extraction Kit is not suitable for any organism that has a cell wall, including algae, plants, yeast, or bacteria.
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.

Can frozen tissues be processed with the Mem-PER Plus Membrane Protein Extraction Kit?

Yes, frozen tissues can be processed with the Mem-PER Plus Membrane Protein Extraction Kit.
How does the Mem-PER Plus Membrane Protein Extraction Kit work?

The kit uses a mild detergent-based selective extraction protocol to extract membrane proteins. First, the cells are permeabilized with a mild detergent to allow the release of soluble cytosolic proteins. A second detergent then solubilizes membrane proteins. Extraction efficiencies will vary depending on the number of times the integral membrane protein(s) of interest spans the lipid bilayer. Membrane proteins with at least 1-2 transmembrane domains are typically extracted with an efficiency of up to 90%. Cross-contamination of cytosolic proteins into the membrane fraction is usually less than 10%.
Can Mem-PER Plus Membrane Protein Extraction Kit be used to extract protein from organelle membranes?

Yes, membrane proteins from various organelles will fractionate with the membrane fraction. We have evaluated COX-IV (mitochondrial membrane protein) and Calnexin (ER membrane protein) which fractionated with the membrane fraction.

What is the expected protein yield after extraction with Mem-PER Plus Membrane Protein Extraction Kit?

Different cell lines will give different yields. Some cells may be more difficult to permeabilize and some proteins may be more difficult to solubilize. For example: Jurkat cell yields from 5 million cells are around 100 µg of membrane protein and 500 µg of cytosolic protein while HeLa cell yields from 5 million cells are around 450 µg of membrane protein and 750 µg of cytosolic protein.

What species/cell lines have been tested with Mem-PER Plus Membrane Protein Extraction Kit?

The Mem-PER Plus Membrane Protein Extraction Kit has been evaluated for use with cultured mammalian cells and tissues (both hard and soft). Cell lines tested include HeLa, HEK293, A431, PC3, LNCaP, A549, HEPG2, and Jurkat cells.

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.
How can I selectively isolate membrane proteins?

Certain detergents can be used to selectively extract and isolate membrane (hydrophobic) proteins from cytosolic (hydrophilic) proteins. The Mem-PER Plus Membrane Protein Extraction Kit (Cat. No. 89842) enriches for integral membrane proteins and membrane-associated proteins from cultured mammalian cells or tissue via selective solubilization using a mild detergent-based protocol. The use of selective detergent extraction eliminates the hassle of phase separation based on hydrophobicity, allowing better reproducibility and higher throughput. The cells are first permeabilized with a mild detergent, allowing the release of soluble cytosolic proteins, after which a second detergent solubilizes membrane proteins.

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