Application: LFA
I was able to visualize the gold nanoparticles attach itself to where I pipetted my secondary antibody (control line). However, I am not able to visualize the nanoparticles attach itself to the antigen as clearly (test line). How can I fix this problem? I am using the max concentration of the antigen (1 mg/mL) as well so that should not be the problem. I have also confirmed that the gold nanoparticles are conjugated with the antibody and that the corresponding antigen is pipetted onto the nitrocellulose membrane. I would greatly appreciate any assistance. Thank you in advance :)
L
If your observations are not flawed by any trivial errors I would suggest that the gold nanoparticle by coincidence is bound to the recognition site of the antibody which now no longer binds efficiently to the antigen but is still recognised by the secondary antibody. You could try to change your conjugation method.
Dear brust,
I also encouter this problem, so in your opnion, what elso method could be used?
Because standard method of passive absoprtion is almost the same.How should I avoid binding to the recognition site?
B.C.
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