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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