Hello Histonetters, I am currently trying to work out a new DAB protocol to be used with IHC on FFPE sections.
I am testing three different protocols, and am having limited success. The first protocol requires me to make a DAB stock solution of 10mg in 1 ml dH20, which is then filtered and frozen immediately. This protocol suggests using DAB tetrahydrochloride XH20, but I am using the free base of DAB as that is what was on hand. The free base does not want to go into solution, but has produced some reaction in my IHC. My second protocol asks for 40mg DAB in 1.5 ml Tris buffer (0.05M, pH 7.6), to make my stock solution. I am again using the free base, and have been stirring since 11:30 am yesterday (MDT) with the result being a chocolate milk-like substance. My third protocol requires me to make up the DAB fresh each time I want to use it. This protocol also asks for the tetrahydrochloride form, but again I used the free base and did get some reaction. Does anyone have thoughts on about how to get the free base to go into solution without oxidizing, or thoughts about free base versus the other form of DAB? Thanks! Keri Keri Colwell Laboratory Technologist | Technologiste de laboratoire TSE and Pathology Lethbridge Laboratory | Laboratoire de Lethbridge Canadian Food Inspection Agency | Agence candienne d'inspection des aliments Township Road 9-1 | Ch de Canton 9-1 Box 640 | CP 640 Lethbridge, AB T1J 3Z4 E-mail | Courriel: [email protected] Telephone | Téléphone: 403-382-5500 Facsimile | Télécopieur: 403-382-5583 Government of Canada | Gouvernement du Canada _______________________________________________ Histonet mailing list [email protected] http://lists.utsouthwestern.edu/mailman/listinfo/histonet
