Dear Reagn-etters,
I post this cry for help on behalf of a friend. Since neither of us
subscribe to this newsgroup, please send any reply to my address, i.e.
lj1 at mrc-lmb.cam.ac.uk. Thank you!
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
Voice: +44.1223.402443
FAX: +44.1223.213556
Telex: 81532
E-Mail: lj1 at mrc-lmb.cam.ac.uk [lj1 at 131.111.84.16]
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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