Some cell surface markers might be degarded by collagenase or dispase during tissue processing, therefore I am looking for alternative protocols. I use collagenase-based mixes to process tumors and retrieve immune cells and further analyze them through flow cytometry. I see Sigma sells collagenase alternative "Accutase": does anyone have experience with that?
Stefano,
I use Accutase on tissue culture cells (stem cells & differentiated cells) when I harvest them to run flow cytometry. Seems to work fine for the surface markers I look at. Supposed to be more gentle compared to trypsin. I have no idea how it would work on tumors though.
Other companies (Corning & MP Biomedicals) sell Accutase besides Sigma by the way.
Good Luck!
Hi, I do not have much experience with tissue dissotiation, but some time ago I bookmarked this:
http://www.worthington-biochem.com/tissueDissociation/default.html
Which I think might help
we normally use a cocktail of collagenase I and IV with Dnase. I saw in literature that my cell marker of interest is affected by any type of collagenase, therefore I am looking for an alternative that does not harm immune cells while dissociating solid tissues
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