Annette, In our hands enzymatic treatment (collagenase/dnase) does not have any significant impact on cell surface markers, including ones that are notoriously protease-sensitive (e.g. Lectin receptors). One thing you should be wary of is tryptic contamination of your collagenase batch, but then again, you prepare a digestion medium with your enzymes in complete medium, including serum, which will inhibit any residual tryptic activity present. So all that youčre left with is specific matrix-degrading activity. You may want to check specific cocktails of enzymes for specific organ dissociations. Worthington Biochemicals (where we buy our collagenase) has an extensive manual and background info on the subject ( http://www.tissuedissociation.com/ ). Feel free to mail for more details. Karim -- Karim Y. Vermaelen, MD Dept. of Respiratory Diseases Ghent University Hospital 7K12ie De Pintelaan 185 Ghent B-9000 Belgium tel: +32 9 240 2605 fax: +32 9 240 2625 On 15/7/2002 19:22, "Annette Byrne" <AByrne@pcyc.com> wrote: > Good morning Flow-ers ! > > > Just a quick query on single cell suspension prep for Flow: Am about to try > making cell suspensions from tumor and aortic tissue for Flow with fluorescent > membrane antibodies. My question is whether it's better to mechanically > disaggregate tissue or enzymatically (collagenase etc). Does enzymatic > tretatment have any negative effect on cell surface antigens ? > > Many thanks as always, > > Annette > Annette Byrne PhD > Scientist > Pharmacyclics > 995 E Arques Ave > Sunnyvale > CA 94085-4521 > USA > > Tel: 001-408-3283640 > Fax: 001-408-3283689 > e mail : AByrne@pcyc.com > www.pharmacyclics.com > >
This archive was generated by hypermail 2b29 : Sun Jan 05 2003 - 19:26:15 EST