[Histonet] Re: BrdU staining

From:Chengkang ZHANG

Hi Stephen,

I assume that OE means Olfactory Epithelium. To eliminate the autofluorescence, you can try to incubate your sections in 5mM CuSO4 (50mM NH4OAc, pH5.0), after last step of your immunofluorescence staining, for 30 min to reduce lipofuscin-like autofluorescence, then wash in ddH2O briefly and in PBS for 5 min. This helps a lot in my hands. You can refer this paper for further reference.

Stephen A. Schnell, William A. Staines and Martin W. Wessendorf. "Reduction of Lipofuscin-like Autofluorescence in Fluorescently Labeled Tissue", The journal of Histochemistry and Cytechemistry, 47(6), 719-730, 1999.

Good luck


> Message: 1
> Date: Sun, 14 Jan 2007 13:37:04 -0600
> From: "Stephen Clark" 
> Subject: [Histonet] DAB brdU protocol
> To: 
> Message-ID: 
> Content-Type: text/plain;	charset="iso-8859-1"
> Does anyone have a protocol for DAB staining, specifically for mouse OE sections at about 10 or 18um using brdU?  I had been using fluorescent staining procedures, but i wanted to try something non-flourescent to elimate any problems with autoflourescence.  I have tried it a few times already with a rough protocol supplied by a colleague with little success.

Chengkang Zhang Ph.D.
Room 357, MedSurge II
19182 Jamboree Rd,
Department of Pharmacology
University of California, Irvine
Irvine, CA 92697-4625
Email:  chengkaz@uci.edu
Tel:    (949)-824-1902 (lab)
Fax:	(949)-824-4855

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