[Histonet] Polymer detection kit on rat tissue

[Liz Chlipala]

Anita
 
Since you are working on rat tissue, you have to be careful on what detection systems you use.  You will need two different detection systems for your work.  One for the mouse monoclonal antibodies and one for the rabbit polyclonal.  If you want to use a polymer based detection system then here are some basic rules you will need to follow.
 
1.  For blocking its probably easiest to use a serum free protein block - many vendors have them.
 
2.  For the rabbit polyclonal antibody you can use any vendors anti-rabbit polymer, but you can NOT purchase a dual link polymer (one that will detect both mouse and rabbit primary antibodies) that will lead to background staining
 
2.  For the mouse antibody on rat tissue if you are dead set on using a polymer then you would need to purchase one that is specifically designed to be used for mouse antibodies on rat tissue.  Biocare Medical and MaxVision have them, there may be others out there these are the two that we have used in our lab.
 
3.  Polymer based reagents are nice but they are expensive.  I don't want you to get the empression that I'm knocking polymer based reagents, we use them in our lab all of the time.  But there are other methods or detection systmes available for use on animal tissues if you are on a budget.  The key to good success with animal detection systems if you are going to use a secondary antibody rather than a polymer is to make sure that the secondary antibody that you purchase has been cross absorbed to the species you are working with. You must also check the spec sheets they will normally give you a cross reactivity list of tissues that have been tested.
 
Good Luck
 
Liz

________________________________

From: histonet-bounces <@t> lists.utsouthwestern.edu on behalf of amitapandey <@t> torrentpharma.com
Sent: Thu 2/24/2011 3:11 AM
To: histonet <@t> lists.utsouthwestern.edu; histonet-bounces <@t> lists.utsouthwestern.edu
Subject: [Histonet] Polymer detection kit on rat tissue



Dear histonetters,

I am lost in data sheets of  IHC detection kits . Please help me  to
finalize one good detection kit for my purpose.

I am planning to do immunohistochemistry on paraffin embedded rat kidney
tissues for mouse monoclonal and rabbit polyclonal antibody (not  double
labelling).
I am looking on Dako envision, vector immpress, Novolink , the company
like Biocare/ Max bio vision suggest the Rat specific detection kit....Got
confused...


Also send me  your opinion on ABC detection versus polymer detection
system.


Thanks in advance for your view.

Amita
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