[Histonet] RE: Controls for nickel enhanced DAB

[David Finkelstein]

Hi Bob,
Cobalt Nickel DAB is my favourite was of intensifying IHC [2].  This is then
counter stained with Neutral red to give good quality images.

Leaving out the primary antibody is often an adequate control especially if
you can identify the antigen in at a specific location in the brain.  

"I'm staining for receptors in mouse CNS, and don't see any staining in 
control sections and some diffuse staining of varying intensity in the 
experimental tissue." It sounds like you have to try playing with the level
of fixation.  Receptors are very difficult to visualise and in my previous
studies with the Dopamine transporter fixation is crucial [1].

It is very difficult to design experiments with quantitative  densotometric
IHC analysis whereas it is, it is relatively easy to do stereological
studies 


1.	Parish, C.L., D.I. Finkelstein, J. Drago, E. Borrelli, and M.K.
Horne, The role of dopamine receptors in regulating the size of axonal
arbors. Journal of Neuroscience, 2001. 21(14): p. 5147-57.
2.	Adams, J.C., Heavy metal intensification of DAB-based HRP reaction
product. Journal of Histochemistry and Cytochemistry, Journal of
Histochemistry and Cytochemistry, 1981. 29: p. 775.

Assoc. Professor David Finkelstein,
The Mental Health Research Institute of Victoria,
155 Oak Street, Parkville, Victoria 3052 
AUSTRALIA 
Mobile: 0409171227
Tel: +61 (03) 9388 1633 
Fax: +61 (03) 9387 5061
dfinkelstein <@t> mhri.edu.au

d