Overview

  • Product name
    Plasma Membrane Protein Extraction Kit
  • Assay type
    Direct
  • 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.

  • Tested applications
    Suitable for: Purificationmore details

Properties

  • Storage instructions
    Store at -20°C. Please refer to protocols.
  • Components Identifier 50 tests
    Homogenize Buffer NM 1 x 100ml
    Lower Phase Solution WM 1 x 20ml
    Protease Inhibitor Cocktail (Lyophilised) Red 1 vial
    Upper Phase Solution NM 1 x 20ml
  • Research areas
  • Relevance
    Membrane 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.

Applications

Our Abpromise guarantee covers the use of ab65400 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Purification Use at an assay dependent concentration.

Images

  • Membrane protein assay - ab65400
    Membrane protein assay - ab65400Image from Choveau F.S et al., PLoS One 10(12), Fig 2C. Doi: 10.1371/journal.pone.0145367.

    Cell membrane protein was collected using plasma membrane protein extraction kit (ab65400). The upper panels show the plasma membrane myc-tagged protein; the middle panels show the cytosolic myc-tagged proteins and the lower panels show the total myc-tagged protein from CHO (Chinese Hamster ovary) whole-cell lysates.

Protocols

References

This product has been referenced in:
  • Wang H  et al. Targeted production of reactive oxygen species in mitochondria to overcome cancer drug resistance. Nat Commun 9:562 (2018). Read more (PubMed: 29422620) »
  • Das S  et al. Intranasally delivered small interfering RNA-mediated suppression of scavenger receptor Mac-1 attenuates microglial phenotype switching and working memory impairment following hypoxia. Neuropharmacology 137:240-255 (2018). Read more (PubMed: 29738851) »

See all 37 Publications for this product

Publishing research using ab65400? Please let us know so that we can cite the reference in this datasheet.

Customer reviews and Q&As

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88 reviews or Q&A

10
Votes: Highest First

I just wanted to give an update on the replacement kit. I have had much much better success with recovering the plasma membrane fraction after we spoke on the phone. I still do not get 100% purity, but the contamination is not too bad. I never have con...

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Thank you for the update. I am glad to hear the replacement kit worked out better for you. Eliminating those extra steps will probably, as you found, give purer fractions but possibly at the cost of lower yield.

I am not sure why tubulin was g...

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confusion about the protocol. why does the upper reagent need to be taken out the bottle before adding into the tube again (step 3 to 5 in the protocol)

Merci pour votre requête. J'ai contacté le laboratoire et ils m'ont confirmé que cette étape est pour équilibrer la "upper phase" solution. Il ne faux donc pas prendre la solution directement de la bouteille....

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Can this kit be used to purify just the plasma membrane, rather than the plasma membrane proteins?

This kit may be useful for extracting and retainingthe plasma membrane fraction without the plasma membrane proteins but it has not been specifically designed for this purpose. Please let us know how things go if you do choose to use ab65400 for this e...

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1.       I would like to isolate a 4 transmembrane protein (504 amino acid) from plasma membrane using ab65400 Plasma Membrane Protein Extraction Kit (attached please find manual).


2.       It is very important for us to understand t...

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I can confirm that the kit uses an aqueous polymer two-phase system to separate plasma membranes. Most specifically it uses the PEG/ Dextran system and the plasma membrane tend to concentrate in the upper phase.


I am sorry we do not have ...

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1. Can this kit be used with smaller cell numbers, such as 105 or 106? If so, what protocol changes are required, if any?
2. Is there a limit to the number of transmembrane domains a protein can have and still be solubilized with this kit?

1. The smaller cell number can decrease yield. Hence it is at the user's discretion to reduce cell numbers. For some cells, based on size and density of expression of membrane proteins, smaller cell number can still yield enough membrane proteins. If c...

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1. I would like to isolate cell membrane protein and membrane associated protein but not mitochondrial membrane protein from fibroblasts.
2. Also It is important to keep protein with native form. Does the protease inhibitor cocktail also work...

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1. This kit isolates the plasma membrane associated proteins as well as transmembrane proteins on the plasma membrane. Organelle membrane proteins are not isolated. If total membrane proteins were desired, you would get all membrane proteins ...

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I was looking at the plasma membrane protein extraction kit. I'm looking to analysis protein expression of a gene of interest on the surface of the membrane vs internalized gene. My question would be if this kit, would give me the sufficient results to...

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Yes, this will give you two sub-cellular fractions: plasma membrane and cytosolic. So you will be able to demonstrate the relative amounts of your protein of interest in each fraction using an assay such as western blotting, assuming you have an antibo...

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Do you know if this kit extract intact-native proteins?

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

The homogenization step is defective, I have homogenized cells in Dounce homogenizer for 50 times, when I did not get a nice result, I put the sample in liquid nitrogen, then tried to grind the sample with the Dounce homogenizer pistol. Do you have any...

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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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I have tried the plasma membrane extraction kit again with no success! I still did not get any pellet at the end, and here are the details:

I am using T47D breast cancer cells.

I have used around 6x 108 cells; wet weight was around 1....

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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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1-10 of 88 Abreviews or Q&A

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

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