发信人: leohawk (leohawk), 信区: Biology
标 题: Re: Anybody know about immunofluorescence
发信站: The unknown SPACE (Fri Sep 13 15:13:56 2002), 转信


involves fixation of sample, primary antibodies, secondary antibodies(with
fluorescence).

use primary antibody from different source, say, Protein A with rat derived,
while protein B with mouse derived, better be monoclonal, if not,
pre-absorption using irrevalent sample from the same organism is usually
required.

The secondary antibodies are all commercially available, like anti-Rat,
anti-mouse... they are fluorescence labeled, there is a lot of choice...
I personally think cy3 gives the strongest signal. In your case, make sure
use 2' antibodies that give different fluorescence.

usually, pre-absorption of the 2' antibody is also required for good results

A good confocal or deconvolution miscroscope is necessary.


【 在 iminusc (newdriver) 的大作中提到: 】
: I'm trying to detect co-localization of 2 proteins in the brain and someone
: suggest to use immunofluorescence. I never do it before and wonder if anybody
: who know it give me some advice. Thanks


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