RE: need buffer "recipe"

From:Roy Ellis

Andrea
I'm not sure that this is the one you want but I found it at
http://www2.swmed.edu/rogersmr/perfusates.htm

Regards
Roy Ellis
mailto:roy.ellis@imvs.sa.gov.au


Krebs-Henseleit (KH) Bicarbonate Buffer

Chemical Conc.  (mM) Amount per Liter

NaHCO3  25.0mM  2.10 g/L
NaCl 118mM 6.90 g/L
KCl 4.7mM 0. 35 g/L
MgSO4  (anhy) 1.2mM 0.145 g/L
NaH2PO4 1.2mM 0.145 g/L
CaCl2 1.2mM 0.175 g/L

Start bubbling the high quality de-ionized water with 95% O2 / 5% CO2.
Add the NaHCO3 first, and the CaCl2 last.  This will allow enough time for
the CO2 to bring the pH of the water down (CaCl2 precipitates at a high pH).
Add the desired water-soluble substrate.
The pH of KH buffer will be 7.4 only if the perfusate is at a temperature of
37o C and gassed with 95% O2 / 5% CO2.





> -----Original Message-----
> From: Andrea Grantham [mailto:algranth@u.arizona.edu]
> Sent: Thursday, 11 October 2001 1:33
> To: histonet@pathology.swmed.edu
> Subject: need buffer "recipe"
>
>
> Does anybody have a "recipe" to make up Krebb's buffer. I've already
> consulted my collection of reference books and where I find many types of
> buffers none go by this name.
>
> Thanks!!
>
> Andi
> .....................................................................
> : Andrea Grantham, HT(ASCP)     Dept. of Cell Biology & Anatomy     :
> : Sr. Research Specialist       University of Arizona               :
> : (office:  AHSC 4212)          P.O. Box 245044                     :
> : (voice:  520-626-4415)        Tucson, AZ  85724-5044    USA       :
> : (FAX:  520-626-2097)          (email:  algranth@u.arizona.edu)       :
> :...................................................................:
>            http://www.cba.arizona.edu/histology-lab.html
>
>





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