I want to cross-link collagen and from literature I found that the most common used buffer is MES and I do not have it yet. Is PBS a better option or distilled water? I know that EDC hydrolyses in water and so its cross-linking efficiency is diminished.
High concentration of EDC(＞＝50ｍM) can effectively crosslink collagen even though in EDC/water solution. So MES is unnecessary if you use high concentration of EDC. EDC will hydrolyse in water, so you need fresh EDC solution for cross-linking. Using ethanol/water mixture as solvent for EDC would be better, because it prevent dissolving of collagen during crosslinking.
I have already tried with ethanol/water and it seems to have a better cross-linking degree (but I will see if it dissolves in PBS after freeze drying). It happened to me before when I used only water (with EDC and NHS).
Depending on what type of material you are working with (collagen gel/espun fibers) you can crosslink collagen dehydrothermally (DHT) by putting your sample into a vacuum oven overnight at 140 C. This only works with solid samples though.
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