I work on Troponin I, I use 19C7 as a capture, and M18 for detection both at 1ug/mL, I need the assay to detect down to 0.1 ng/mL, and I'm just getting signal at 50 ng/mL. I use 2% BSA for block. I tried G-protein coated plates, different concentrations of the antibodies, even the wash, I wash originally washing all with tween, then tried washing all with PBS, and the last step only with tween, which obviously didn't work, and I even switched Avidin to Streptavidin, using tghe HRP detection system. Can you help, please? Thanks
Have you tried independently varying the concentrations of the 2 antibodies and the amount of time for each binding step?
I did try different concentration combinations, but haven't tried the binding times.
You might also be able to find a more sensitive detection reagent. If you have access to a luminescence plate reader, you could try this one, for example:
https://www.lifetechnologies.com/order/catalog/product/37070
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