I will need some assistance here. I need someone help me to put real numbers in place of the variables in the following protocol. You don't need to know anything about this particular method or science, just how to plug in numbers. I'll be isolating adipocytes from adipose tissue using collagenase. I need you to tell me how much collagenase I will need along with how much DMEM and other reagents.
http://www.serva.de/www_root/documents/Prot_adipocytes_pre_human.pdf
I will be digesting about a cup of adipose tissue at a time.
I dont see whats wrong with that protocol....all the neccersary info is in there...the only thing you have to do is to calculate/measure the volume of starting material.
I know, I just need help doing it. I've never worked with collagenase before and the units are in mg and everything else is in ml.
So if you need 0.3 Units/ml and you have for example, 3 ml, that is 0.9 Units or 0.3 x 3. If the collagenase is 0.1 unit/ mg, this comes to 9 mg of collagenase you need for 3 ml. So hopefully, the collagenase probably comes in mg, and you just weigh out what you need.
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