Next generation RIPA buffer: a new kit for Lipid Raft

UltraRIPA kit tebu-bio funakoshi

RIPA buffer is one of the most useful protein extraction buffers and maintains most native structures of proteins. Nevertheless, RIPA buffer is not convenient enough to extract membrane proteins and membrane-associated proteins concentrated in lipid raft.

Lipid raft is a highly unique microdomain on the lipid bilayer which contains special lipids, cholesterol and functional proteins. In this layer, lipid raft-enriched proteins are usually insoluble in mild detergent buffers such as 1% Triton X-100 and RIPA buffer. Consequently, it was, up to now, difficult to analyze the functions of this kind of proteins.

Whato do then?

UltraRIPA kit for Lipid Raft provides a solution to extract and analyze membrane proteins or membrane-associated proteins enriched in lipid rafts with native structure and function.

UltraRIPA kit tebu-bio funakoshi

Features of UltraRIPA kit

  • Extract membrane / membrane associated proteins enriched in lipid rafts, with native structure and fully retained function
  • Easy and simple procedure : only a centrifuge is required
  • Only two components: A buffer and B buffer

Example Data

Extraction of RIPA-insoluble proteinsExtraction of RIPA-insoluble proteins WB

Extraction of RIPA-insoluble proteins

Left : Quantification of extracted total proteins by BCA protein assay. UltraRIPA kit could constantly extract over 70% of RIPA-insoluble proteins from the mouse brain tissue.
Right : Western blotting of lipid raft markers. Some lipid raft markers among proteins or a ganglioside extracted were dramatically increased in RIPA-insoluble fraction by UltraRIPA kit.

 

Enzyme Activity AssayTotal protein phosphatase activity

Enzyme activity assay of proteins extracted by UltraRIPA kit

Left : Lactate Dehydrogenase activity of proteins extracted by using 1% Triton X-100, A and B buffer. Equivalent enzyme activity are obtained when using A and B buffer.
Right : Total protein phosphatase activity. Protein extracts from RIPA-insoluble fraction of the mouse whole brain by UltraRIPA kit B-buffer, RIPA, and 2% SDS buffer were applied to total protein phosphatase assay. Proteins extracted by UltraRIPA kit had high phosphatase activity.

Convinced? Try it now!

Related products:

Learn more about Montana Molecular kits, designed to help you in GPCR mediated signal detection.

Don’t hesitate to contact our specialists to know more and discover a wide range of tools for the search of transmembrane and signaling pathways proteins.

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Written by Elodie Monin
Community Manager