I develop a lateral flow assay using PS microspheres conjugate. I realise my tests strips by strips and the assembly is handmade.
I pretreat the sample pad with PBS Tw20 Sucrose buffer and dry it 2 hours at 37°C in a dry box. After the pretreatment of conjugate pad with 10mM phosphate Tw20 Sucrose buffer and drying, I dip the conjugate pad in a conjugate solution and dry it 2 hours at 37°C in a dry box.
A part of the conjugate don't flow across the membrane in the presence of a liquid sample. It is partially blocked in entrance of nitrocellulose membrane (aggregation) but a part arrives to migrate because my control line appears. However if I place a new strip (without conjugate on conjugate pad) in a conjugate solution, the conjugate migrates correctly across the nitrocellulose membrane, without be blocked.
I had been facing similar problem.
Using Triton x-100 to 0.25% in your chase buffer would help you reduce or prevent aggregation
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology
- Immunology Techniques