[Histonet] Anti-DIG peroxidase problem

Tony Henwood AnthonyH <@t> chw.edu.au
Tue Dec 6 14:51:55 CST 2005

Try using a FITC labelled probe, followed by a mouse or rabbit anti-FITC.
Then use your usual immunoperoxidase demonstration technique (ABC or polymer - I use a peroxidase-labelled polymer)


Tony Henwood JP, MSc, BAppSc, GradDipSysAnalys, CT(ASC)
Laboratory Manager & Senior Scientist
The Children's Hospital at Westmead,
Locked Bag 4001, Westmead, 2145, AUSTRALIA.
Tel: 612 9845 3306
Fax: 612 9845 3318

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Mikael Niku
Sent: Tuesday, 6 December 2005 6:09 PM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Anti-DIG peroxidase problem

Dear Histonetters,

I'm trying to set up tyramide amplification for my in situ 
hybridizations with digoxigenin probes.
Unfortunately it seems as if the anti-DIG-POD by Roche doesn't work as 
well as the anti-DIG-AP I'm normally using.
Basically, I get a weaker signal with anti-DIG-POD + tyramide 
amplification than with unamplified anti-DIG-AP.

I have also tried biotinylated probes to avoid using anti-DIG-POD 
altogether, but so far I haven't got any signal at all with them.

Any ideas, anyone?

With best regards,
Mikael Niku


  Mikael Niku             URL: www.helsinki.fi/~mniku/
  University of Helsinki  Dept. Basic Veterinary Sciences

  - Mitäkö mieltä olen länsimaisesta sivistyksestä?
  Minusta se olisi erinomainen ajatus!

                                          - Gandhi


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