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Jeremy -

Possibly a stupid question, but your description is somewhat ambiguous in 
that respect: Have you ever done a direct comparison, i.e. used the same 
primary abs for direct and indirect labeling?

Beat

At 16:36 30/07/02, you wrote:
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>http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1nfocal
>
>Until recently I had always been under the impression that indirect
>immunocytochemical methods were more sensitive than direct conjugates.
>However, in the past couple of weeks I've had much better results with three
>separate unrelated directly conjugated monoclonals (anti-Integrin alphaE,
>-IgE and -beta catenin) than any other primary/secondary system I've ever
>used.
>
>Indeed, I go so far as to say that 1 µg/ml of direct conjugate yield vastly
>superior signal and negligible background when compared to 1°/2°,
>biotin/streptavivdin or even TSA based methods. I really don't understand
>why I have to use higher concentrations of 1° antibody for indirect methods
>(usually 2-10 µg/ml).
>
>I'd also like to take the opportunity to moan about MP refusal to licence
>the Alexa dye series to other manufacturers.
>
>Has anyone tried their monoclonal labelling kits out?
>
>Perhaps its just me, so I thought I'd open up a discussion on the subject.
>
>Jeremy
>
>****************************************
>Dr. Jeremy Brown
>Research Fellow
>Department of Veterinary Clinical Studies
>Royal (Dick) School of Veterinary Studies
>Easter Bush Veterinary Centre
>Easter Bush
>Roslin
>Midlothian
>EH25 9RG
>Scotland
>UK
>
>http://www.image-analysis.co.uk
>****************************************
>