How to extract intracellular lipids in lipidomics research
The process of extracting intracellular lipids usually includes the following steps:
1. Cell Collection:
Collect the cultured cells by washing and centrifugation.
2. Cell Lysis:
Use a lysis buffer (such as a detergent-containing salt solution) to disrupt the cell membrane and release the cellular contents. This step ensures the release of lipids.
3. Lipid Extraction:
The most commonly used methods are the Bligh and Dyer method or the Folch method. These methods typically involve the use of organic solvents (such as chloroform, methanol, and water).
- Folch Method: Add chloroform and methanol (ratio of 2:1) to the lysed sample. Shake thoroughly to ensure effective lipid extraction.
- Bligh and Dyer Method: Add methanol, chloroform, and water (ratio of 2:2:1.8) to the sample. Shake thoroughly.
4. Centrifugal Separation:
After appropriate shaking and mixing, the sample is centrifuged to separate into two layers: the upper layer is the aqueous phase (containing non-lipid components), and the lower layer is the chloroform phase (containing lipids).
5. Lipid Collection:
Carefully collect the lower layer, which contains the extracted lipids.
6. Solvent Evaporation:
Use nitrogen drying or a vacuum centrifugal evaporator to remove the solvent, obtaining a pure lipid sample.
BiotechPack, A Biopharmaceutical Characterization and Multi-Omics Mass Spectrometry (MS) Services Provider
Related Services:
Lipidomics MALDI Imaging Services
Mammalian Untargeted Lipidomics
How to order?
