Has anyone worked with desmosine elisa kit before? I'm trying to determine desmosine content in aorta tissue from rats. according to the kit protocol, for tissues i have to mince the tissue and follow the elisa procedure. so we know that desmosine is crosslinked in tissue, does mincing break it down? basically what i'm measuring ? free desmosine or desmosine from elastin. let's say if i have a tissue with less elastin content (diseased tissue) shall i get a higher value or less compared to a healthy tissue?
If you want to measure the amount of tissue elastin following demosine content determination you are expected to completely release the total desmosine present as one of the specific crosslinking compounds of insoluble elastin. Usually this step is accomplished by total acid hydrolysis in 6 N HCl at 108 °C up to 72 hour followed by acid removal by rotary evaporation. If you are not following this procedure you are likely to measure possible free desmosine unlikely to be present in tissues.
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