[Histonet] RE: Double immunostaining with a rabbit and rat
C.M. van der Loos
c.m.vanderloos <@t> amc.uva.nl
Tue Apr 3 02:20:51 CDT 2007
I cannot recommend you this G/2 kit as this is sequential as well as
multi-species based. A home-brew solution is more appropriate here:
Rt CD34 + Rb casp-3 (cocktail)
Goat anti-rat/HRP (mouse absorbed)* + goat anti-rabbit/AP polymer **
Develop AP activity in red (Dako LPR)
Develop HRP activity in brown (DAB+ from any source)
* = see Jackson or Rockland
** = see LabVision
Lots of success with staining!
Chris van der Loos, PhD
Dept. of Pathology
Academic Medical Center M2-230
NL-1105 AZ Amsterdam
Date: Mon, 02 Apr 2007 14:40:59 +1000
From: Cathy Malcontenti-Wilson <cmalc <@t> unimelb.edu.au>
Subject: [Histonet] Double immunostaining with a rabbit and rat
To: histonet <@t> lists.utsouthwestern.edu
We wish to do a double immunostain for CD34 and active caspase 3 on
paraformaldehyde fixed mouse tissues. The cd34 primary is rat
anti cd34 The caspase 3 primary is a rabbit polyclonal antibody.
We would like to use the Dako Envision G/2 kit for double staining,
this kit is for mouse or rabbit primaries only. When we do a single
for cd34, we use a Rabbit anti Rat IgG as a linking antibody between
primary and secondary antibody (Dako Envision plus kit for rabbit
primaries) and this works well on our tissues in our experience.
My question is: Can I add this linking antibody step to the method and
still use the Envision G/2 kit?
Does anyone have ! any exper
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