I'm trying to measure OVA-specific IgE with a DIY ELISA, with an OVA coating. The issue I am facing is that since in the model I am using OVA-specific IgGs are produced in much higher concentrations than IgE, low dilutions of the serum get lower signal than higher dilutions. This reduces my signal significantly compared to my standard, which works great.
Would pre-incubating the sera on anti-IgG-coated plates help reduce that signal? And if so, has anybody done that before?
Thanks!
İsmail Emir Akyildiz
Deplete the serum using protein A to get rid of IgG prior to the ELISA app. Collect the flow-through fraction resulting from the spin column or cartridge and dilute with the proper buffer and volume...
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