[Histonet] Re: renin

[Jose Luis Palazon Fernandez]

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" <mauger <@t> email.chop.edu> 
>Subject: renin 
>To: jluis.palazon <@t> icman.csic.es, histonet <@t> lists.utsouthwestern.edu, lpwenk <@t> sbcglobal.net
>
> Hi Lee and Pegy,
> 
> Do you know of a source for +renin control?
> 
> 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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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