Protein Lipid Overlay (PLO) Assay


Protein Lipid Overlay (PLO) Assay Platform

In addition to serving as a physical membrane bilayer to produce cell compartments, lipids also play an important role in signal transduction, material transport, morphological changes, and cell division through specific lipid-protein interactions. The systematic study of the interaction between lipids and proteins can reveal the functions of lipids in cells and help us understand lipids more comprehensively.

A variety of techniques for studying lipid-protein interactions have been developed, such as nuclear magnetic resonance spectroscopy (NMR), surface plasmon resonance (SPR), isothermal titration calorimetry (ITC), liposome sedimentation assay, and native mass spectrometry and so on. Protein lipid overlay (PLO) analysis can identify lipid ligands that interact with lipid-binding proteins, and the assay can also provide qualitative information on the relative affinity with which a protein binds to a lipid. This method only requires a few micrograms of protein, and is fast and inexpensive. But the technology still has limitations that must be overcome. To explore more unknown functions of lipids, new and/or improved techniques are essential for detecting lipid-protein interactions.

 The key steps of the protein lipid overlay assayFigure 1. The key steps of the protein lipid overlay assay (Dowler, S.; et al. 2002)


In the classic PLO assay, lipids are spotted on the membrane, and then a series of operations are performed, including blocking with blocking proteins, incubating with the epitope-tagged target protein, incubating with the primary and secondary antibodies, and detection. Compared with the classic PLO assay, the PLO developed by Creative Proteomics is more efficient and more sensitive. In addition to significantly improving the signal of the interaction between lipids and their interacting proteins, our PLO method may also be suitable for studying lipid-protein interactions that cannot be detected by the traditional PLO method. Therefore, our PLO method has a wider application range, which can help customers expand their research on lipids and their binding proteins, and reveal new functions of lipids in different organisms.

We provide a high-quality one-stop service, including all steps in the PLO experiment process, such as:

  • Lipid packaging and dissolution
  • Lipid spotting and drying
  • Blocking and incubation
  • Washing
  • Incubation with primary antibody, wash
  • Incubation with secondary antibody, wash
  • Detection

Customers can choose different technology platforms according to project requirements, or contact us directly for consultation, and our expert team will provide you with customized experimental procedures.


  • Lipid-protein interaction study
  • Lipid function study
  • Lipid-binding protein research
  • Unknown interaction discovery

Creative Proteomics is an international biotechnology company dedicated to research in molecular interactions and other related fields. After continuous improvement, our protein lipid overlay assay platform has greater sensitivity and can analyze interactions that cannot be analyzed by classic PLO. Our one-stop service aims to save customers time and money.


  1. Dowler, S.; et al. Protein lipid overlay assay. Science Signaling. 2002(129).
  2. Han, X.L.; et al. An improved protein lipid overlay assay for studying lipid-protein interactions. Plant Methods. 2020, 16(1).
* This service is for RESEARCH USE ONLY, not intended for any clinical use.