Re: periodate-lysine-paraformaldehyde fixative

From:Paul Klosen <>

At 12:24 20/06/01 -0700, Ford, Judi {Path~Palo Alto} a écrit:
>Hi everyone!
>I was wondering if anyone has a detailed recipe for PLP fixative.  One of my
>clients wants to use this fixative for
>their immuno work-up and we don't have anything on it in our lab.

Dissolve 7.3 g of lysine monohydrochloride in 200 ml of ddH2O. Adjust pH to=20
7.4 with 0.1 M Na2HPO4  (Na2HPO4 . 2 H2O 17.8 g/l) NOT phosphate buffer 
!!!. Complete volume to 400 ml with 0.1 M phosphate BUFFER (!!) pH 7.4. 
This lysine-phosphate buffer keeps for 2-3 days in the refrigerator, but 
can be frozen in aliquots for longer storage.
Just before use, mix 375 ml lysine-phosphate buffer with 100 ml 20% 
Formaldehyde and top to 500 ml with ddH2O. Add 1.06 g NaIO4 and mix well. 
The PLP fixative must be used within maximum 2 hours.

Final concentrations: Lysine 75 mM, Formaldehyde 4%, Sodium periodate 10 mM

The original PLP fixative by Nakane & co used 2% Formaldehyde and a 
variable amount of periodate. This is the mix that works best for us for 
both ICC and ISH.

                                                                        (o -) O
Paul Klosen, PhD
CNRS UMR 7518 Neurobiologie des Fonctions Rythmiques et Saisonnieres
Universite Louis Pasteur  12, rue de l'Universite
F-67000 Strasbourg, FRANCE
Tel.  Fax.

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