how should one perform an ELISA to detect antigen-specific IgE?
what blocking and buffer do you use?
do you dilute the serum?
what detection method are you using?
Dear Oliver,
If you want, you can glance at the ELISA method that we described in our paper Medicine (Baltimore) 2015 Oct;94(40):e1699. PMID: 26448021.
I hope it's useful.
Dear Maria, many thanks!
I read your paper. very nice study! did you play around with the ELISA conditions? e.g. test different dilutions of the sera?
which BSA did you use for blocking? how high where wells without patients serum? with blocking buffer and the HRP alone?
i know those SouthernBiotech antibodies very well, i also tested them. I think Anisakis leads to very high levels of IgE maybe. Did you test allergens under the same conditions maybe? do you know the total IgE of the patients? would be nice if this correlates somehow with the specific IgE.
Article Previous Exposure to the Fish Parasite Anisakis as a Potenti...
Hi Oliver,
we perform a sandwich ELISA: Article Displaying Fel d1 on virus-like particles prevents reactogen...
If you detect nothing, you can try to use serum at 1:4 or even 1:2 dilution.
Cheers
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