Hi all



is renin antibody an expensive one?. O just want to know if juxtaglmerular cells are present in an aglomerular fish species I am studying. I only would use it for some 4 or 5 slides and I dont know if it is worth while to buy the antibody As my lab is not specialized in inmunostaining. 

 thanks in advance for replys



José Luis



El dia 12/07/2005 18:38 usted envio el siguiente mensaje:

>Date: 12 de Julio de 2005 18:38:41

>From: "Joanne Mauger"  

>Subject: renin 

>To: jluis.palazon@icman.csic.es, histonet@lists.utsouthwestern.edu, lpwenk@sbcglobal.net

>

> Hi Lee and Pegy,

> 

> Do you know of a source for +renin control?

> 

> Thanks,

> Jo Mauger

> 

> >>> "Lee & Peggy Wenk"  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=0D
> 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. =0D
> 

> 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=0D
> 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@lists.utsouthwestern.edu 

> [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Jose Luis=0D
> Palazon Fernandez

> Sent: Wednesday, June 29, 2005 9:01 AM

> To: histonet@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@icman.csic.es  

> 

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Universidad de Oriente-Isla Margarita-Venezuela

actualmente en: Instituto de Ciencias Marinas de Andalucia

Puerto Real, Cádiz, España.

email: jluis.palazon@icman.csic.es

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