The source of my Glucagon was Human Glucagon from European Pharmacopoeia(EP) Reference Standard from Sigma-Aldrich; I used a few different plate conditions with (1) Standard Nunc Plates, (2) Nunc Plates with a Poly-Lysine coating and (3) the Greiner Non-binding plates.
Experimental Conditions were 0.6mg/ml Glucagon dissolved in 3.2mM HCl, 0.9%w/v NaCl with 50 microMolar Tht. There was 5s of automixing before each fluorescence measurement.
The second plates scenario worked the best in terms of getting something close to a typical sigoimdal curve but in the other scenarios I was getting some weird behaviour. Has anyone seen anything similar? Attached is a pdf of my overlaid-raw Tht assay data.
Any help would be appreciated