Ian,
        are you sure that these figures aren't reversed? As you know, in most
instances, fluorescence is much more easily detected than using markers with
brightfield.
A concentration of 1:20 would also suggest that the antibody is either not very
concentrated or binds weakly.
If you using this antibody would suggest that as you are probably using pieces
of tissue for confocal, that you consider a dilution with longer binding times
and perhaps changing the solution once, as this has been shown to increase the
total amount of antibody bound (with some antibodies).
Is your tissue fixed or fresh?
Barry

Ian Montgomery wrote:

>         Just bought , from Chemicon, a mouse anti-human collagen type VI
> monoclonal antibody (Cat.No. MAB3303). If anyone is using it, what dilution
> are you using, a ball park figure will do. I asked Chemicon and they say
> start at 1:250 for brightfield and 1:20 for fluorescence. I'll be using the
> antibody for a confocal study so must use fluorescence. By the time I do a
> dilution trial starting at the suggested 1:20 and with only 50ul of neat
> antibody the study is going to cost a fortune. Any ideas on dilution will
> be most welcome.
> Ian.
>
> Dr. Ian Montgomery,
> West Medical Building,
> University of Glasgow,
> Glasgow,
> G12 8QQ,
> Scotland.
> Tel: 0141 339 8855 Extn. 6602.
> Fax: 0141 330 4100.
> e-mail: ian.montgomery@bio.gla.ac.uk

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