[Histonet] RE: Manual IHC for Melan-A

[Goins, Tresa]

We do manual staining for everything - here is the abbreviated version for Melan A -

Thermo Scientific MS799
Clone A103; Mouse MAb Isotype IgG1
	
Protocol:

1.	Dewax and hydrate sections
2.	Retrieve with HIER - EDTA pH 8 20 min  
3.	Block with NGS for 15 min
4.	Treat with primary Ab for 30 min
5.	Apply Link #1 - biotinylated α-mouse
6.	Apply Link #2 - streptavidin enzyme complex
7.	Rinse with water and apply chromogen 
8.	Rinse with dH2O 
9.	Counterstain, dry and coverslip

Results:
	Staining is cytoplasmic
	Granular cytoplasmic staining of adrenal cortical cells; minimal or no staining of kidney
	Equivalent results using canine and feline tissue during validation

Tresa
	


-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Michelle M. Mackinnon
Sent: Thursday, November 08, 2012 9:00 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Manual IHC for Melan-A

Has anyone done manual IHC for Melan-A?  If yes, who help set up the manual protocol?
Thanks,
Michelle

Michelle M MacKinnon HT, (ASCP)
Histology Supervisor
Dermatology Consultants. PA
1215 Town Centre Drive, Suite 200
Eagan, MN 55123
Phone: 651-255-3462
Fax: 651-255-3461

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