RE: immuno sensitivity

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From:"Connolly, Brett" <brett_connolly@merck.com>
To:"'Judy Trogadis'" <judy@playfair.utoronto.ca>, "'histonet'" <histonet@Pathology.swmed.edu>
Reply-To:
Date:Tue, 18 May 1999 14:00:51 -0400
Content-Type:text/plain

Judy,

Without a doubt you should try tyramide signal amplification (TSA). This is
a perfect example of what this technology was developed for. You can use
fluorescent or chromagenic detection. Check out the NEN website for details

http://www.nenlifesci.com/ti_link1.htm

Brett


Brett M. Connolly, Ph.D.
Senior Research Biologist
Dept. of Human Genetics
Merck Research Laboratories
WP26A-3000
PO Box 4
West Point, PA 19486
tel. 215-652-2501
FAX 215-652-2075

> ----------
> From: 	Judy Trogadis[SMTP:judy@playfair.utoronto.ca]
> Sent: 	Tuesday, May 18, 1999 11:59 AM
> To: 	HistoNet@Pathology.swmed.edu
> Subject: 	immuno sensitivity
> 
> Histonetters,
> 
> I am trying to detect a protein in brain tissue using immunocytochemistry.
> From northern and western blots we know that the RNA is made and is 
> translated into protein, now we want a more precise localization - it
> should
> be nuclear but we are not sure in which cell type(s) it is expressed.
> 
> I have used a biotinylated IgG followed by the ABC kit from Vector which
> is 
> a peroxidase reaction using DAB. No protein was detected although grossly,
> the tissue had more of a yellowish tint compared to negative controls.
> Also,
> we have a positive control which uses the same tissue preparation, i.e.
> fixation, cryosectioning, etc. One current theory is that the protein is
> 
> present in many cell types but at a low level of expression.
> 
> What would you say is the most sensitive immuno-detection method
> available?
> Would fluorescence be better?
> 
> Thanks in advance
> judy
> 
> 
> Judy Trogadis
> Eye Research Institute and
> University of Toronto
> Toronto Hospital, Western Div.
> 399 Bathurst  St.
> Toronto, Canada M5T 2S8
> 
> phone: 416-603-5088
> Fax:   416-603-5126
> email: judy@playfair.utoronto.ca
> 
> 
> 



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