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Buffers that are good for cell culture. - need buffering capacity around pH 5-7 (Feb/05/2009 )

I'm doing an assay which requires me to keep cells grown in a 6well plate at low pH (around pH 5.5), however, the pH-adjusted PBS I'm using has caused my cell monolayer to peel off the floor of my 6well plate after incubation of >30 min. , I'm looking for a buffer that has good buffering around pH 5.5 - 7.0, at the same time, I need something that is cell culture friendly, anybody know of something of that sort?

Any help or advice would be appreciated,



What cell line are you growing?

-Minnie Mouse-

In general, cells in culture do not "like" being in PBS for long periods of time. Adhesion usually requires Mg and Ca, do you include these in the PBS? If the assay permits I would have used a balanced salt solution such as Earle´s or Hank`s. If possible include some protein (serum or BSA for example). As for buffering I would suggest 10-20 mM Hepes . This is commonly used in culture medium such as Iscove`s modified MEM.

Hope this helps


HEPES is an excellent choice for normal pH range. For your particular pH condition, I would suggest MES, perhaps?