Measuring rat plasma samples with ELISA using commercial kits (Spi-Bio A05201) with prion standards from Switzerland. But our calculated results are in the ~100's ng/mL (including control groups). But looking through publications, other groups have listed control values as typically
You might look up papers from Ed Hoover's lab (Colorado State University).
Thanks, we figured it out. Simple math problem.
Any troubleshooting tips would be appreciated
hi Nathan,
try to make a 2, 5 or if highly concentrated 10 fold dilution of your plamsa sample. (Better would be to treat them with PNGase F).
For control positiove, use a recombinant prion protein with known concentratio, you can make dilutions to have a series of concentrations too.
Treat both of them with proteinase K if you're looking for pathological form of the protein.
Load equal quantities of the samples on gel for western blot
You can have an idea of the exact quantity
For more precise calculation, you may consult Dron et al. 2011 JBC
for your convenience, I put here the concerned part,
"Quantification of Deglycosylated PrPSc Species
Samples were treated with PNGase F according to the manufacturer's instructions (New England Biolabs) and quantification of full-length and C2 PrPSc bands was determined by GeneTools software after acquisition of chemiluminescent signals with a GeneGnome digital imager (Syngene)."
Good luck
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