I have a researcher interested in quantifying erythroid progenitors using ter119 and CD44 as markers in early stage mouse embryos that have been dissociated with collagenase. Here's my questions: 1) Is there a better way to dissociate the cells and reduce clumping? (metalloproteinases?) 2) Should we resuspend in a special buffer (PBS & EGTA)? 3) Can we leave the cells in the collagenase to reduce clumping or will the cells become too fragile? 4) Will the collagenase cleave off the epitopes for ter119 and CD44? 5) Does anyone have suggestions for better markers? -Derek Schulze Flow Cytometry and Confocal Microscopy Cancer Research Labs, 353 Botterell Hall Queen's University Kingston, ON, K7L 3N6 Canada
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