I want to know how many molecules I can localize and quantitate in an section in one shot? What are the things I should take care while buying the antibodies.
You visit www.abcam.com orhttp://www.antibodyresource.com/onlinecomp.html
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Thanks for replying my query but I am not finding the solution. My problem is that I want to localize 3 molecules in a single slide for confocal microscopy, so I want to know how to select these three primary and secondary antibodies? My experimental animal is rat and I want to do it in brain sections. Please help.
Each of your primary antibodies should be raised in a different species.
target 1 Primary Ab - Rabbit Polyclonal
Target 2 Primary Ab - Mouse monoclonal
target 3 Primary antibody - raised in chicken
then the secondary antibodies conjugated to each color fluorphore that you wish to use should be be for anti-rabbit, anti mouse and anti chicken in this example. It is best to use preconjugated secondaries in this case to minimize target cross-detection
And try to make sure that spectra for your secondary antibodies has as little degree of overlap as possible so you can get some clean images.