Help in IHC

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20101975's picture
Help in IHC

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.

MUTHUMARIAPPAN Murugan's picture
Arvind Singh Pundir
Arvind Singh Pundir's picture

i would suggest you a truly valuable link where you can search all your needs about antibody search and reviews, techniques and applications
do come back with all your querries
Good luck !

20101975's picture
20101975's picture
Thanks for replying my query

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.

The FFM's picture
Each of your primary

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

heehawmcduff's picture
And try to make sure that

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.