According to the experimental procedures provided in several reference , I attempted to remove endogenous uPAR from the cell surface using an acidic solution. The protocol involved washing the cells with 50 mM glycine-HCl and 100 mM NaCl (pH 3.0) for 1 minute (repeated three times) to remove surface-bound endogenous uPA. Subsequently, the cells were neutralized with 0.5 M Hepes and 0.1 M NaCl (pH 7.5) for 10 minutes on ice. However, after returning the cells to the cell culture incubator following these treatments, their morphology changed drastically within 10 minutes. The cells appeared to be excessively digested, exhibiting shrinkage, rounding, and clustering, resembling the effects of over-digestion by trypsin.
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