Overview

  • Product namePlasma Membrane Protein Extraction Kit
  • Assay typeDirect
  • Product overview

    The Plasma Membrane Protein Extraction Kit provides optimized buffers and reagents for effective extraction of membrane proteins from mammalian tissues and cells. Unlike other available procedures that can only extract the total cellular membrane proteins (combinations of plasma and organelle membrane proteins), this kit was designed to not only extract the total cellular membrane proteins, but also purify the plasma membrane proteins specifically. The procedure offers consistent yield and high purity (over 90%). Membrane proteins prepared using the kit can be utilized in a variety of applications, such as Western blotting, 2-D gels, and enzyme analyses, etc. The entire procedure takes less than 1 hour.

Properties

  • RelevanceMembrane Protein Extraction is a method for extraction of membrane proteins from mammalian tissues and cells. Many procedures extract total cellular membrane proteins which is a combination of plasma and organelle membrane proteins.

Protocols

References for Plasma Membrane Protein Extraction Kit (ab65400)

This product has been referenced in:
  • Yang H  et al. Inhibition of PP2A activity confers a TRAIL-sensitive phenotype during malignant transformation. Mol Cancer Res 12:217-27 (2014). Read more (PubMed: 24296757) »
  • Wang Z  et al. p21-activated kinase 1 (PAK1) can promote ERK activation in a kinase-independent manner. J Biol Chem 288:20093-9 (2013). Read more (PubMed: 23653349) »

See all 11 Publications for this product

Product Wall

There is no detergent in any component of this kit.

The proteins isolated when using this kit should be in their native conformation.

The frozen tissue cannot be pulverized with the Dounce homogenizer. This has to be done with a pulverizer or tissue grinder. Once ground, the buffer can be used to homogenize the bits in a Dounce homogenizer.

Fresh tissue can be homogenized di...

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You are obtaining a huge pellet after step 4. This is the low speed centrifugation at 700g and pellets down unbroken cells, nuclei and debris. Getting a huge pellet at this step is indicative of incomplete homogenization. The steps are correct but...

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I would not recommend the syringe method over the Dounce homogenizer method. The Dounce method is the most gentle yet thorough way to make sure the membranes are in solution homogenously. The proteins might be damaged due to shear forces if drawn ...

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The idea is to homogenize in as much buffer as required, not too much, not too little. Our lab uses 1-2 mL of wash and homogenization buffer with 5x10^8 cells homogenizing with a dounce homogenizer for their preps to validate this kit with good results...

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Most unbound lipids are removed during the process of purifying plasma membrane proteins. This kit could work for bacteria but the bacterial cell wall has to be broken before the plasma membrane can be accessed. This can be done either by using lysozym...

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Unfortunately, I have to confirm you that the homogenization buffer do contains Tris. Therefore, if the Tris is affecting the activity of the enzyme you are studying I won't suggest you to use this kit.


If you stop after section A and use the total cellular membrane protein pellet, the majority of the unstimulated receptor will be most likely in this fraction. But depending on how big the membrane vesicles are, it could be in the cytosolic fracti...

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We have used Cox 4 and Cadherin as membrane protein markers in our preparations using ab65400 and have seen no cross-contamination between cytosol and membrane fractions.

The expected membrane protein recovery is 70-100 ug/50 millio...

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Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"