Simple extraction protocol for membrane proteins

 Membrane proteins are often quite challenging to extract and below you will find components of a simple extraction buffer which can be applied. 
  • 4 M urea
  • 10 mM DTT
  • Triton X-100

Material was frozen in liquid nitrogen. Thoroughly and promptly grounded material was kept on ice for 30 minutes followed by subsequent centrifugation at 10 000 xg for 4 minutes. 

Protein determination can be done using Pierce™BCA Protein Assay Reagent (Thermo Fisher Scientific) which is compatible with urea and detergent present in extraction buffer. 

This protocol has been successfully applied for protein extraction and detection with Agrisera antibodies to the following membrane proteins: ATG8, NBR1 and PDAT1.

References:
Klińska-Bąchor et. al (2023). 

Klińska-Bąchor et. al (2025). 		 

Simple extraction protocol for plant membrane proteins


 

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