Preparation of FAT conjugate for Mycoplasma HELP !

FRANCOIS DREYER francie at vetlab8.agric.za
Thu Jul 20 05:01:49 EST 1995


I hope I am posting this to the right place.

We started a mycoplasma section at our lab.
I now want to prepare  FAT (Fluorescence Antibody) conjugates to ID
the isolates.

I have a method I can use but there is one part where I am stuck. It reads
as follows :

" Place the conjugate in a dialysis *bag*
  Let it concentrate in a 10% solotion of Polyethylene glycol 4000
  Dissolve the PEG 4000 in phosphate buffer.
  Wash the outside of the *bag* with distiled water after concentration. " 
  etc.

I found a company that told me it is not *bags* but membrane tubings.
The problem is that there is a lot of different ones depending, as I 
understand, on the Molecular weight of the sample you are using.
I now want to know what size? membrane to use.

The institute that is doing the FAT ID for us at the moment is using 
conjugates that has been prepared by a lady that does not work there anymore
and I can't trace her. Nowbody at this institute knows how to prepare these 
conjugates.

If anybody out there can help me or give any references that I can use
please reply to my e-mail.


Thank you all

Francois Dreyer  <francie at vetlab8.agric.za>


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