Establish of an indirect ELISA in one month possibleHiston
Posted 03 May 2013 - 08:55 AM
during my thesis i want to quantify the if Histon 3 or 4 gets more methylated\acetylated in a specific tissue of mice living in different conditions. Therefore we have several Antibodys for both, ELISA and Western-Blot. First we tried Westernblot and it works fine but we need to much of the sample (10 ug per blott) and the material is very limited (down to 60 ug per sample). Furthermore stripping didnt work because we lose to much of the proteins or the signal is still there. Therefore we think about to establish an indirect elisa in our lab but the problem is i have only two months to go for my diploma thesis. No one done ELISA before in our lab so iam on my own.
I want to ask if is hard to establish an indirect ELISA (the antibodys are working very well for western blot) and how much protein from a whole tissue is per detection necessary ? Abcam recommend 1ug per well, is this right ? Is ELISA suitable to detect and quantify very fine methylation\acetylation changes of Histons between different samples ?
I hope somebody can answer me this basic questions. Thanks !!!
Posted 06 May 2013 - 07:12 PM
Posted 07 May 2013 - 05:25 AM