Re: [Histonet] nickel enhanced DAB procedure for rat brain

From:Rene J Buesa

Neil: 
  Nickel "enhancement" (a name used to develop  a blue-purplish chromogen colour) uses a 16.2% aq.sol. of nickel sulfate hexahydrated in the following proportion:
  a- DAB ---- 6 mg
  b- PBS ---- 10 mL
  c- 3% hydrogen peroxide --- 160ÁL
  d- 16.2% nickel sulfate aq.sol. ---- 120 ÁL
   
  Adjust the recipe for your 10 mg DAB tablets.
  Hope this will help you.
  RenÚ J.

Neil Fournier  wrote:
  I was wondering if anyone has experience using nickel enhanced DAB? One of our protocols suggested using nickel enhanced DAB. We use 10 mg DAB tablets available from Sigma Chemicals to make up our DAB substrate solution. We dissolve our DAB in .1 M phosphate buffer (pH 7.4). 

Does anyone have suggestions for making up a nickel-DAB solution. If so, what is the typical concentration, recipe, and procedures for making up this solution? I also realize that some individuals dissolve the nickel ammonium sulphate in acetate buffer. Is this necessary? And if so, what is the rationale for this. 

Appreciate the advice,

Neil
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