Coating Buffer for Sandwich ELISA
Posted 27 January 2005 - 12:37 PM
I've been trying to coat a custom peptide onto Immulon plates because I am trying to develop an assay. Sometimes I get signal, sometimes I don't.
I'm wondering whether it will be because the coating of the "sample" is not done well?
I use 1x PBS + 0.02% Tween-20 for coating.
Any other coating buffers out there? How would I know which type of buffers are best for my peptide, sample, antibody, etc?
Posted 28 January 2005 - 03:51 AM
I used a carbonate-coatingbuffer:
0.1 M Na2CO3
0.1 M NaHCO3
coated 100µl/ well of a 96 well plate (medium binding capacity, but maxisorp plates worked equally well) at 4°C over night.
worked for all kinds of proteins. if it doesn't work with your protein, may changing the pH wpoulod be a good idea.
Posted 04 February 2005 - 04:05 PM
Does this mean that I should only block once in the protocol and just use phosphate buffer to dilute any antibodies I need to use in the protocol?