Dear All,
I need to coat my hydrogel ( Methylcellulose) coated dishes with Collagen, the protocol that I have from the company says that I should dissolve Collagen in Acetic Acid, but Acetic Acid degrades my hydrogel. On the other hand I'm not sure if the collagen will dissolve evenly if I change the pH? Do you have any suggestions?
I think you use can use water to dissolve collagen by heating. Then you can avoid toxicity and pH problems.
It depends from the type of collagen you have. If it is a partially hydrolysed sample, as often is, it should dissolve in hot water and settle again into a hydrogel on cooling down to room temperature or lower (5-10°C). The settling process is usually slow, might even take hours, so you have all the time to coat your plates. Just try first on a piece of glass to establish the best operating conditions.
normally you use an acetic acid buffer and low temperature (4°C) to dissolve higher Mw collagen, protocols are described in literature. Mostly it is done by steering overnight in a fridge, as it dissolves very slowly. Once it is dissolved you can dilute the stock solution further and bring the pH to 7 (eg. use phosphate buffer, for example PBS). it depends on how you want to use the samples afterwards but the easiest way of coating them is dip-coating with the collagen solution and use the hydrophobic-hydrophobic attraction. Good luck.
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