Krebs buffer cloudy

S.E. seedelm at yahoo.com
Wed May 12 10:01:12 EST 2004


I am making up Krebs buffer for an isolated-perfused heart
preparation.  The recipe for the buffer is as follows: NaCl 118mM,
NaHCO3 25mM, KCl 4.8mM, KH2PO4 1.2mM, MgSO4 1.2mM, Glucose 11mM
CaCl2(2H2O) 1.5mM and Sodium Pyruvate.

I was told to dissolve the chemicals in order and make sure that each
item is dissolved completely before I add the next chemical or else
the buffer will become cloudy.  Unfortunately, even if I follow those
directions, the buffer will still become cloudy as soon as I add the
CaCl2 quite frequently.

Another researcher suggested to bubble the solution with 95%CO2/5%O2
for 5 minutes before adding the CaCl2, but that didn't solve the
problem.

I'd really like to be able to be able make to make this buffer
regularly without having to throw out the first 3 preparations because
they cloud up as soon as add the CaCl.....

Any suggestions? 

Thanks is advance, 
Stephanie



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