I am developing a competitive lateral flow device for the determination of antibiotics in milk. I am struggling with uneven staining of the test line. The test I developed has 4 test lines and I have a problem with only the third test line. I tried to change the concentration, pH, detergents, alcohols, and sugars in the test-line dispensing buffer and no improvement was achieved. The other test lines and control lines don't have such a problem. The protein I dispense on the third test line is BSA-chloramphenicol conjugate.
Could you advise on the problem?
As a sample, I use milk that doesn't contain any antibiotics.
I use liquid gold conjugate for testing.
I attached the photo of the same sample run in multiplicate (23 total runs) at the same time.
try mixing in a bit of methanol, only a very low concentration to the mixture you add to draw the line
Thanks for the answer.
We tried using 5% methanol in the dosing buffer, but there was no improvement.
Is a bit far fetched, but is it possible that this could be a problem of the machine you use to draw the line? could the tip be creating the problem by applying the solution with interruptions?
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