Hi
Enzyme digestion is an antigen retrieval system too, so guess you mean heat
or enzyme first - on bone marrow trephine biopsies for Igs, we used heat
retrieval followed by a brief enzyme. Haven't heard of it the other way
round.
Cheers

-----Original Message-----
From: histonet-request@lists.utsouthwestern.edu
[mailto:histonet-request@lists.utsouthwestern.edu] 
Sent: 02 May 2005 18:04
To: histonet@lists.utsouthwestern.edu
Subject: Histonet Digest, Vol 18, Issue 2

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Today's Topics:

   1. fungus control blocks (Ernestine Middleton)
   2. Ki67 - kidney. (christian@eisbo.dk)
   3. antigen retrieval/enzyme digestion (Till, Renee)
   4. Re: Ki67 - kidney. (Kelly D Mcqueeney)
   5. Normal donkey serum block solution (=?gb2312?q?=CC=EC=20=D0=C1?=)


----------------------------------------------------------------------

Message: 1
Date: Sun, 1 May 2005 15:54:30 -0400 (EDT)
From: Ernestine Middleton 
Subject: [Histonet] fungus control blocks
To: histonet@pathology.swmed.edu
Message-ID: <20050501195430.97482.qmail@web51502.mail.yahoo.com>
Content-Type: text/plain; charset=us-ascii

Hi;
Sorry, my e-mail address is ernestinemiddleton@yahoo.ca.
 
Ernestine.



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Message: 2
Date: Mon, 02 May 2005 10:31:07 +0200
From: "christian@eisbo.dk" 
Subject: [Histonet] Ki67 - kidney.
To: histonet@lists.utsouthwestern.edu
	
Message-ID: 
Content-Type: text/plain; charset=windows-1252

Hey users of histonet.
I am trying to stain rat kidneys for Ki67!
I have bought the recomended antibody. But this one is for human!
Do anyone know if this can be used since i dont have a outcome!
Do anyone have a protocol that works?
Christian Eisbo



------------------------------

Message: 3
Date: Mon, 2 May 2005 07:55:02 -0500
From: "Till, Renee" 
Subject: [Histonet] antigen retrieval/enzyme digestion
To: histonet@lists.utsouthwestern.edu
Message-ID:
	
Content-Type: text/plain; charset=us-ascii

When  trying both on the same slides, which would go first, the antigen
retrieval or the enzyme digestion?

 

Renee' Till, HT

 

Arkansas Childrens Nutrition Center

1120 Marshall

Little Rock, AR 

72202

 

(501) 364-2774

 


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Message: 4
Date: Mon, 02 May 2005 09:26:49 -0400
From: Kelly D Mcqueeney 
Subject: Re: [Histonet] Ki67 - kidney.
To: "christian@eisbo.dk" 
Cc: "histonet@lists.utsouthwestern.edu"
	
Message-ID: <42762A99.7010505@bms.com>
Content-Type: text/plain; format=flowed; charset=windows-1252

Hi Christian,
I have used rabbit polyclonal human Ki67 antibody on rat kidney and it
worked beautifully.  Make sure you use a wash buffer with tween-20. 
Simply follow the protocol recommended by Abcam (ab833). I used heat antigen
retrieval in citrate buffer (Dako), 3% BSA (only) as a block and 
Envision+ rabbit kit (Dako) rather than ABC. The incubation time is O/N
at 4C.

Let me know if you need more detailed information. You can also use Ki67 
antibody from Santa Cruz.

Good luck,
Kelly

christian@eisbo.dk wrote:

>Hey users of histonet.
>I am trying to stain rat kidneys for Ki67!
>I have bought the recomended antibody. But this one is for human!
>Do anyone know if this can be used since i dont have a outcome!
>Do anyone have a protocol that works?
>Christian Eisbo
>
>_______________________________________________
>Histonet mailing list
>Histonet@lists.utsouthwestern.edu
>http://lists.utsouthwestern.edu/mailman/listinfo/histonet
>  
>



------------------------------

Message: 5
Date: Mon, 2 May 2005 21:30:08 +0800 (CST)
From: =?gb2312?q?=CC=EC=20=D0=C1?= 
Subject: [Histonet] Normal donkey serum block solution
To: histonet@pathology.swmed.edu
Message-ID: <20050502133008.26004.qmail@web15507.mail.cnb.yahoo.com>
Content-Type: text/plain; charset=gb2312

Hello,all,
 
I am doing double-immunofluorescence. Both the hosts of my secondary
antibodies are donkey, so I plan to use normal donkey serum for blocking,
but I do not know how to make up this solution. 
I have ordered donkey serum from sigma company, but the specification
doesnot show how to make up it. so if you have done it, I hope that you can
do me a favor. 
In addition, if the hosts of my secondary antiboies are different, one is
horse, the other is donkey, which solution do I use for blocking? Thank you!
 
Best wishes!
 
Guofeng
 
Giessen
 
 



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