I want to isolate connexin from rat brain in milligram quantity. I have been suggested to first isolate the crude membrane faction and then only connexin can be obtained. At present I am trying to implement protocol proposed by Suski et al. 2014 for isolation of crude membrane fraction. But I don't have much idea for further purification of connexin.
Can someone suggest any other approach for isolation of connexin from rat brain ?
Thanks a lot.
http://www.nature.com/nprot/journal/v9/n2/full/nprot.2014.016.html
Hope this helps!! http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1991.tb21075.x/pdf
You can please follow the links below:-
1. http://www.ncbi.nlm.nih.gov/pubmed/20396696
2. http://onlinelibrary.wiley.com/doi/10.1111/j.1432-1033.1991.tb21075.x/pdf
3. http://connection.ebscohost.com/c/articles/48126375/isolation-extracellular-recombinant-fragment-rat-connexin-43
Well, I'm the first author of the affinity purification paper. Try to make an affinity matrix with the antibody against the C term of Cx43 supplied by sigma and try to include a gel filtration step to remove the lipids before affinity Chromatography but you will need a LOT of rat brain if you want mg....
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