[Histonet] LFB with Vector ABC kits

[Geoff McAuliffe]

Hi Nick:

    You may have to 'do the experiment' yourself to answer the stain 
compatibility question. Vector's "Intense" reagent makes DAB a very nice 
gold color, should look nice with LFB. I have had excellent results with 
"Intense". I suspect that it is a gold cholride-based intensifer.
    As for lithium carbonate, I as pretty sure that it is the carbonate 
(ie alkalinity) you want, not the lithium, so try something with one 
(more?) CO3 ion. Years ago one of the grad students in the anatomy dept. 
at U. of Louisville med school did some testing on lithium versus other 
alkalies, I don't remember exactly what she found.

Geoff

Donin, Nick (NIH/NCI) wrote:

>I am trying to stain using LFB in combination with the Vector ABC Staining
>kits.  For the Vector Kit staining, I am using a biotin-conjugated secondary
>antibody, which then conjugates with an avidin-peroxidase molecule which
>reacts with a DAB substrate.  Does anyone know if these two methods are
>compatible?  If so, is it preferable to do either one first?  Additionally,
>my lab has run out of Lithium Carbonate for the LFB protocol, can I
>substitute with Lithium Chloride?  I'm trying to stain formalin fixed,
>paraffin embedded mouse brain.  Thanks for your help.  Email me any
>questions or suggestions you have.  Thanks.   
>
> 
>
>Nick Donin
>
>CRTA
>
>Neuro-Oncology Branch
>
>National Cancer Institute
>
>National Institutes of Health
>
>9000 Rockville Pike
>
>Building 35, Room 2B-203
>
>Bethesda, MD 20892
>
> 
>
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Geoff McAuliffe, Ph.D.
Neuroscience and Cell Biology
Robert Wood Johnson Medical School
675 Hoes Lane, Piscataway, NJ 08854
voice: (732)-235-4583; fax: -4029 
mcauliff <@t> umdnj.edu
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