I don't have a reference at the moment. But I use DIG-labeled oligos
for ISH. The problem with small probes or low-copy sequences is, that
you have to use a good amplification system.
In our hands a good starting point is the following:
- Anti-DIG antibody from mouse
- Anti-mouse-DIG antibody
- Anti-DIG-antibody conjugated with either FITC or AP or similar
Boehringer Mannheim calls this combination 'DIG amplification system'.
>In article <3264B583.44D8 at alf.biochem.mpg.de>, Gabriele Kerber
><kerber at alf.biochem.mpg.de> wrote:
># Ms. K.K. Martin wrote:
># > I have used both 35S and DIG for ISHH. I far prefer DIG and will never,
># > ever use 35S again. DIG is easier, safer, faster (results in hours rather
># > than days or weeks you get good spatial localisation and, best
># > of all, you avoid all that nasty paperwork associated with keeping track
># > of your radioactive waste. Boehringer Mannheim have very helpful manuals
># > for using DIG in ISHH. Good luck.
># DIG is wonderful for IHS with riboprobes. But if you want to use
># oligonucleotides, you will probably have a lot of problems with
># DIG. With oligos it4s better to use 35S or even 32P.
>Is there a published side by side comparison of 35-S vs. DIG in the
>literature? I have the impression, from ISHers I've talked to, that 35-S
>ISH is considerably more sensitive than DIG. However, I have no reference
>for this claim.
>Gene S. Huh, Ph.D.
>Department of Molecular and Cell Biology
>Life Sciences Addition, Room 221
>University of California at Berkeley
>Berkeley, California 94720-3200