rambis at iastate.edu (Paul L Pearson) wrote:
>>I am currently looking into doing double-labelling of pig brain tissue
>for neuropeptides using two rabbit antibodies and a two fluorescence
>detection system. I am considering using unlabelled fab fragments to
>hide the first primary.
>>>If anyone has experience (either good or bad) with this technique, I
>would appreciate references and/or advice that could make this job a
>little easier. Thanks.
>>Please send responses to: rambis at cvm01.vm.iastate.edu>
I do not have a direct experience with this type of double labelling experiments, but a
colleague (which has left the scientific area) developed a method during which uses a
biotinylated and a unlabelled primary antiserum. Thze biotinylated antiserum was labelled with
flourescence labelled avidin whereas the other antiserum was labelled with an ordinary
flourescence labelled secondary antiserum. The Method was described in:
Würden S., Homberg U., (1993) A simple method for immunofluorescent double staining with
primary antisera from the same species.. J. Histochem. Cytochem. 41:627-630
Tho Co Author is now at the University of Regenburg in Germany, but I do not have his email
The labelling of the antiserum with biotin is described in:
A laboratory manual
E. (?) Harlow, D. Lane
Cold Spring Harbour Laboratory 1988
The results achived with this method were really impressive.