Trying to make an ELISPOT protocol based on the old paper.
Want to coat my plate before adding cells of interest, secondary enzyme-linked antibody, chromogenic substrate in agar, etc.
If I just evaporate my solution onto the plate will the antibody stick?
Maybe I should opt for maleimide?
You can try using the usual ELISA protocols for coating antibodies. In general, 10 - 20 ug/mL in PBS or 0.1M NaHCO3 for an hour or overnight (tightly covered - do not evaporate!). Follow by blocking. However, different antibodies may require slightly different concentrations and/or buffers.
Dont evaporate really? The antibody will bind the plastic all on its own?
Yes. Immunoglobulins, like a lot of other proteins, have hydrophobic moieties on their surface, which facilitate spontaneous binding to hydrophobic surfaces such virgin polystyrene. This phenomenon was reported initially (I believe) by Catt and Tregear, Science 158:1570-1572, 1967, in developing radioimmunoassay. Later on, Engval and Perlmann (name spelling may not be accurate) used this finding to develop the micro ELISA.
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