I used a carbon conjugated polyclonal antibody to detect plant viruses, however, I observed both bands on the strip. Both antibodies were employed at a dosage of 1mg/ml. What should I do to eliminate the undesirable outcome?
We used a borate buffer containing 8.8 pH, citrus Tristeza virus coat protein gene-specific polyclonal antibody on test line and secondary anti-rabbit Igg polyclonal antibody on the control line
Okay thanks, still not a lot to go on but I'd make the following observations. 1. pH 8.8 is rather high for LF. It may not be relevant to your background issue but as it's unusual you may wish to think about using a lower pH. 2. You don't say what else is in the borate buffer? I'm assuming you have BSA and a detergent? If not, then I would expect you to have NSB issues. 3. Based on your C line reagent, your detector on the T line must be a rabbit Ab to your virus. Again, probably not relevant to your NSB problem, it would be preferable to use monoclonal reagents on the T line, if only for greater consistency in the future (I suppose your detector could be a rabbit mono?). 4. Finally, carbon is very sticky and most people use gold as it's easier to work with.
What I would recommend is that you try PBS with 1mg/ml BSA and 0.1% Tween 20 as a running buffer, or add the BSA and Tween to your borate buffer, if not already present. If you are still having issues after this, it might be worth trying gold in case carbon is the root cause of your problem.
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