Posted 24 March 2005 - 02:47 AM
I want to study histone modifications affecting a particular gene that I'm working with. I guess ChIP is the method to use, but I (or my lab) have no experience in this method. How hard is it to do, how much time should I expect for this kind of experiment? Any advices? Are there any companies that do it for you?
Posted 24 March 2005 - 02:50 PM
Posted 26 March 2005 - 08:13 PM
may the ChIP gods look upon you favourably.
Pcrman is right in saying it is a very challenging technique to master, but once mastered it's very rewarding to see the results.
Upstate have a very good kit, I would also like to add that Abcam are very good with their antibodies, some if not most histone modification antibodies are guaranteed for ChIP.
I know abcam have sourced their H3K9 trimethylated from the Jenuwien Lab, a very good antibody for heterochromatin.
Be aware with the antibodies though, even though they may well be guarnteed for ChIP I have experienced that this is not the case, so don't go out and buy a big batch of a particular antibody until you are sure the darn thing works! Note the bacth number of that antibody that you test and insist on getting that batch as the activity can vary between batches.
good luck with it and may the ChIP-force be with you always!
All comments and communication are my own personal ones, and are not tied to any of my affiliations.
Posted 30 March 2005 - 12:17 AM
I´ve been looking into some antibodies. I suspect that my gene of interest is deacetylated (due to TSA studies) do I have to try H2A, H2B, H3 and H4, or is one of these sites more likely to be acetylated, or is all of them acetylated if one is...
Could you recommend which one to start with?
Posted 30 March 2005 - 12:45 AM
Posted 27 January 2009 - 11:40 AM