immunohistochemistry problem

Peter Ashby p.r.ashby at dundee.MAPS.ac.uk
Wed Aug 16 09:31:03 EST 2000


In article <0FZD00OGHSNOEP at Salicet.ucd.ie>, teresa.lambe at ucd.ie wrote:

> Hi, 
> 
> I'm not sure if this is the correct newsgroup for this question 
> but ...
> 
> I am having problems with high background staining in my 
> immunohistochemistry and am considering buying a kit, 
> which will conjugate my primary antibody to either my 
> secondary or a fluorescent dye such as FITC.

I wouldn't conjugate your secondary and using a labelled secondary will
always boost your signal strength. have you determined where your
background is coming from? Do it again with a) no antibodies, b) primary
only c) secondary only to tell where your problem is. If it is either of
your antibodies you could either titrate it or wash with high salt washes.
 
> Has anyone used any of these kits and if so which have they 
> found to work well ??

I always buy the reagents separately, gives you more control. Which
chromogen are you using? Alk phos, POD or fluorescent?

You could also try asking in sci.bio.immunocytochem it gets little enough
traffic and could do with some.

Peter

-- 
Peter Ashby
Wellcome Trust Biocentre
University of Dundee
Dundee, Scotland
Reverse the spam and remove to email me.






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