H.Veldman-2 at neuro.azu.nl
Tue Sep 12 02:15:10 EST 2000
Jens Rainer Hansen schreef:
> I have coated ELISA plates with Ceramide - 1000 - 100 - 10 and 0 ng/ml (100
> mcl/well) using 2 coating buffres (ph 9.6 and 7.2).
> Hereafter we added a mouse anti ceramide antibody and a biotinylated rabbit
> anti mouse antibody, TMB etc., with no result. All wells gave the same as
> the "0" = bagground.
> Does anybody have experience with coating procedures using ceramide ?
> Jens R.
Since ceramides are not proteins but glycolipids, it comes as no big surprise
that the usual coating buffers do not work.
You could try the method used for coating with the related gangliosides:
Add 200 nanogram of the specific glycolipid, dissolved in 100 microliter of
ethanol to each well and let dry by evaporation.
Proceed with blocking etc. in the usual manner.
Hope this helps.
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