Ligand Acidification by Nonadherent Cells
The goal of methods for ligand acidification by nonadherent cells is to obtain nearly continuous measurements of the pH to which a receptor-bound ligand is exposed after endocytosis. The protocols presented below are based on the method developed by Sipe and Murphy  for adherent (BALB/c3T3) cells and modified by Sipe et al.  for nonadherent (K562) cells. Reviews of acidification of endocytic compartments [3,4] and flow cytometric methods for analysis of endocytosis [5,6] may be consulted for additional background. The protocols below describe analysis of transferrin (Tf) acidification, but they may be easily modified for use with a different ligand.
KeywordsArgon Fluores Fluorescein FITC Transferrin
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- 5.Murphy, R.F, Roederer, M., Sipe, D.M., Cain, C.C., Bowser, R. (1989) Flow Cytometry: Advanced Research and Clinical Applications (Yen, A., ed) Vol. II, pp. 221–254, CRC Press, Boca Raton, FloridaGoogle Scholar