I tried using a GenwayBio ELISA kit (40-288-10066F), but I had no signal at 450 nm for samples, standards, or controls. Because of this, I suspect it may have been the coating process.
I used the following consumables:
1) Immulon 1B Clear Flat-Bottom Polystyrene Plate (ThermoFisher 3355): Adsorption, hydrophobic binding property.
2) 5x ELISA COATING BUFFER Reagent (GenwayBio GWB-Q00204): Diluted to 1X in deionized water. Incubated overnight, protected from light, covered to avoid evaporation.
3) Blocking Buffer For ELISA (GenwayBio GWB-E120C6).
4) TMB 2-Component Microwell Peroxidase Substrate Kit (SeraCare 5120-0047): ELISA was read at 450 nm
5) TMB Stop Solution (SeraCare 5150-0020)
I’d appreciate any thoughts on this issue. Thank you in advance.
Thank you for the informative responses, Monyer and Assel. I will try these more hydrophilic plates.
Jose, could you describe the whole setup of your experiment? What other antibodies have you been using? What were your positive and negative controls?
I seriously doubt that the amount of hydrophilicity of ELISA plates will yield a works / does not work result.
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