Hi, is there anybody who can share a good protocol for cell membrane (transmembrane) protein extraction from zebrafish brain tissue to be assayed by Western.? I tried one with a soft lysis solution (125 mM NaCl, 50 mM Tris and 10 mM EDTA) followed by centrifugation and resuspension in sample buffer of the pellet ,...but it didn't work out well. Any suggestion? Thanks
Good Day. When we approach a new membrane protein for purification we usually use nanodiscs. SMALPs or DIBMALPs to be precise:
https://cube-biotech.com/what-are-nanodiscs#synthetic
Our protein is usually tagged with a Rho1D4-tag on its C-terminus.
One the first try, I would recommend this schreening Kit for SMALPs and DIBMALPs to identify the best performing one.
https://cube-biotech.com/products/nanodisc-products/synthetic-nanodisc-products/diisobutylene-maleic-acid-dibma/822/synthetic-nanodisc-screening-kit-8x2x50-mg?number=18290
Hi Enrico Moro . A quick and easy way could be using Triton X-114 phase separation.
Article Small-scale Triton X-114 Extraction of Hydrophobic Proteins
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