Complement and plastic microtiters

Luca Jovine lj1 at mrc-lmb.cam.ac.uk
Fri Mar 17 07:12:30 EST 1995


Dear Reagn-etters,

I post this cry for help on behalf of a friend:

I am doing some assays that involve normal human AB+ serum. I test the
serum for classical and alternative pathway activity before storing it at -
70 C. When the serum is thawed and the C5a levels measured, they are very
low. However, after incubation at 37 C in sterile freezing vials or 96 well
tissue culture plates the levels of C5a rise to comparable amounts found in
serum activated by immune complexes. Since the experiments are concerned
with the interactions between complement, immune complexes and various
cells, the fact that the plastic itself appears to cause complement
activation could be a problem.
Has anyone else come across a similar problem? Are there any 96 well
sterile plates which are known not to activate serum, or any coatings that
might help?

Thank you very much in advance for all the help! I'll post a summary if
anyone's interested.

Luca





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   Luca Jovine

   Structural Studies Division
   MRC Laboratory of Molecular Biology
   Hills Road
   Cambridge CB2 2QH
   ENGLAND

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           lj200 at cus.cam.ac.uk           [lj200 at 131.111.8.1]
           lj200 at hermes.cam.ac.uk        [lj200 at 131.111.8.34]
           jovinel at imiucca.csi.unimi.it  [jovinel at 159.149.10.1]

   "I'll give you a definitive maybe" (Sam Goldwyn)

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