[Histonet] CD34 for primate tissue

[Eddie Martin]

Hi Cathy,

I haven't heard of the cross reactivity with monkey species using Novocastra CD34. However, novocastra's antibody is a monoclonal antibody. You may get better staining utilizing a polyclonal CD34 instead. Thermo-Fisher and Abcam provide a polyclonal CD34. If you reach out to them, they may be able to provide you with a sample size 0.1 microliter if they have them available. 

I hope his helps!

Best,
Eddie


Sent from my iPhone

> On Jun 30, 2016, at 9:06 AM, Cathy M. Mathis/Comparative Medicine <cmmathis at wakehealth.edu> wrote:
> 
> Rhesus is a species of old world monkey.  There are a couple of other companies that have this same clone and on their datasheets they say it cross reacts with rhesus monkey.  The CD34s (2) that I have do not state on their data sheets that they cross react with anything except human and yet are the same clone as the companies that say it does.  I guess I will just break down and purchase a different clone that states it works in rhesus monkey.
> Cathy
> 
> -----Original Message-----
> From: Eddie Martin [mailto:edmartin26 at gmail.com] 
> Sent: Wednesday, June 29, 2016 8:22 PM
> To: Cathy M. Mathis/Comparative Medicine
> Cc: histonet at lists.utsouthwestern.edu
> Subject: Re: [Histonet] CD34 for primate tissue
> 
> What species is the kidney you are trying to stain with IHC. Because you mentioned that you used normal human kidney and it worked , but Rhesus kidney didn't stain. I'm wondering if the Rhesus kidney is human or another animal species? The Novocastra CD34 (QBEND10) is intended for human tissue. 
> 
> Best,
> Eddie Martin, HTL,QIHC
> 
> Sent from my iPhone
> 
>> On Jun 29, 2016, at 4:43 PM, Cathy M. Mathis/Comparative Medicine <cmmathis at wakehealth.edu> wrote:
>> 
>> 2 more bits of information about my dilemma; I did try staining 
>> without any retrieval - no signal I am using rhesus kidney as a 
>> control (getting no signal), but I also ran some human kidney and got beautiful staining with a 20 minute HIER using a pH 8 EDTA buffer.
>> So I know the antibodies and my protocols work, just not on my rhesus.  Does anyone know of a CD34 that will work in this species?
>> More suggestions please?  Thank you all for your time.
>> Cathy
>> 
>> -----Original Message-----
>> From: Eddie Martin [mailto:edmartin26 at gmail.com]
>> Sent: Wednesday, June 29, 2016 5:34 PM
>> To: Cathy M. Mathis/Comparative Medicine
>> Cc: histonet at lists.utsouthwestern.edu
>> Subject: Re: [Histonet] CD34 for primate tissue
>> 
>> Hi Cathy,
>> 
>> EDTA (Retrieval #2) for 20 minutes on the Bond-Rx should be sufficient. And all that is necessary. Please contact me if you need additional help. 
>> 
>> Best,
>> Eddie Martin, HTL, QIHC
>> Edmartin26 at gmail.com
>> 
>> Sent from my iPhone
>> 
>>> On Jun 29, 2016, at 8:16 AM, Cathy M. Mathis/Comparative Medicine <cmmathis at wakehealth.edu> wrote:
>>> 
>>> Good morning my fellow Histo-netters, Does anyone have experience 
>>> with any CD34 antibody that would work in FFPE rhesus tissues?  A few companies have one with clone QEBend 10 and say that it works but I have had no luck.  I am using the Bond RX polymer system and I've tried all the epitope retrievals from Leica.  Now I am try epitope retrieval offline; pronase, trypsin, pressure cooker with high and low pH solutions.  Still no signal.  Any help would be greatly appreciated.
>>> Cathy
>> 
> 
> 
>