I have been isolating microvesicles from HT29 and MA104 cell lines. However, I am not sure which antibody to use to validate the isolation. I have been reading about selectin, integrin and flotillin-2. Could someone suggest me the one that would be the most appropriate for cell culture derived microvesicles?
Hello Zubaida Marufee Islam
Microvesicles carry membrane-derived receptors, proteins, including cytokines, chemokines, proteins involved in cellular signaling and/or migration, lipids, carbohydrates, and genetic material including mRNA and microRNAs. Their contents depend on the parent cell, the microenvironment and on the triggers preceding their release.
Microvesicles mainly express surface markers like integrin-β, CD40 and selectins from the cells they originate from. These markers could be used to validate the isolation.
However, for HT-29, which is a human colorectal adenocarcinoma cell line with epithelial morphology, tumor-derived MVs (TMVs) with many surface determinants like CD44H (hyaluronian receptors), major histocompatibility complex I (MHC I), extracellular matrix metalloproteinase inducer (EMMPRIN), chemokine receptors (CCR6, CX3CR1), and epithelial cell adhesion molecules could be identified.
For more information you may want to refer to the articles attached below.
Article Interactions of tumour-derived micro(nano)vesicles with huma...
Article Roles of Microvesicles in Tumor Progression and Clinical Applications
Best.
Do you really need to use affinity based isolation or enrichment? There are options to isolate vesicles by harnessing their physical properties...SEC or Density gradient (e.g. optiprep) would also be usefull...
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