I am working on the crystallization of novel proteins. Here I found and Purified FAD-dependent enzyme and I screen the initial 43 kits for my protein but after one week I have not found any crystal in well of kits. In the meantime, I also checked the purity of protein by running gel before and after the crystallization. Here, I am asking that is there any special factors which are involved in the FAD-dependent enzyme crystallization.
First thing protein purity should be above 95%. Many things matter like, Including temperature, pH and surroundings of crystallization lab. More than that protein stability also matters.
Did you add FAD to your sample or make sure there is FAD inside after purification?
Dear Stefan, thank you very much for your answer FAD is already present in my purified enzyme because of the color but one thing I would like to ask how we will find the bound FAD in my enzyme do you know any assay?????..... and my second question is there any assay for unbound FAD????
Well, I guess there is an assay for your enzyme and it should work without adding FAD if it is already present?
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