[Histonet] RE:Renin by IHC

[Joanne Mauger]

Hi Lee and Peggy,

Noting your reference to renin staining, I have been working up 2 renin abs,and my problem is the need for a good positive control.
Does anyone know where I can purchase one? Or does anyone have a renin producing tumor that they are willing to share?

Thanks,
Jo Mauger

>>> "Lee & Peggy Wenk" <lpwenk <@t> sbcglobal.net> 06/30/05 6:26 AM >>>
Just wondering if doing an immunohistochemical stain for renin or cathepsin
D would be easier, and more reliable, than doing the Bowie.

Also, JG granules will stain positive with PAS (periodic acid-Schiff) and
TFT (thioflavin T), though neither of these stains are specific for JG
granules.

I'll cover the basics of the Bowie stain (taken from the Sheehan book), and
you can let me know if you want still want me to send you the procedure. 

Fix for 48 hours in Helly fixative (mercuric chloride, potassium dichromate,
formaldehyde)(Yes, it must be Helly. Bowie will not work with formalin fixed
tissue)
Wash tissue in running water overnight, then process and embed. Cut sections
at 4 um.

Deparafinize slide to water.
Lugolize (de-Zenkerize) with iodine and sodium thiosulfate
Mordant in 2.5% potassium dichromate at 40 degrees C overnight
Rinse in d. water
Stain in Bowie solution overnight(mixture of Biebrich scarlet, ethyl violet
water, filtered, filter paper with dye allowed to dry overnight, dissolving
dry dye in alcohol)
Blot slides
Dip in acetone
Differentiate in xylene:clove oil mixture (hint from me - clove oil makes
the lab smell great, but is very expensive)
Xylene

Why the stain isn't used any more:
- Fixation and staining takes several days (4 by my count). 
- Tissue must be fixed in Helly.
- Mercury and potassium dichromate waste must be correctly disposed
- Expensive clove oil must be bought(100 mL = $60+ US)

Let me know if you still would like the Bowie procedure. 

Peggy Wenk, HTL(ASCP)SLS
William Beaumont Hospital
Royal Oak, MI 48073

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu 
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Jose Luis
Palazon Fernandez
Sent: Wednesday, June 29, 2005 9:01 AM
To: histonet <@t> lists.utsouthwestern.edu 
Subject: [Histonet] Bowies method

Dear List-fellows  Could any of you send me the protocol of Bowie s method
for juxtaglomerular cells in kidney.  thanks in advance  José Luis
Universidad de Oriente-Isla Margarita-Venezuela actualmente en: Instituto de
Ciencias Marinas de Andalucia Puerto Real, Cádiz, España. email:
jluis.palazon <@t> icman.csic.es  

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