ELISA plate coating with plasma membrane vesicles - (Jun/06/2011 )
Has anyone coated their ELISA plates with plasma membrane vesicles? Any suggestions on how this might be done? Thanks!
A quick literature search shows this has been done mainly back in the 80 and 90s in a number of applications. The approach was to use a regular coating approach, either neutral PBS or basic carbonate buffers and low to mid microgram per mL concentration of total protein using maxisorp, polysorp or other vendors' equivalents.
Fixation with low concentration glutaraldehyde (0.5% or less in PBS) may make the vesicles more stable if the antigen still remains accessible, but this might make blocking more challenging. Active coating plates (see Pierce) may also be helpful.
Start simple before adding potentially unhelpful complexity.