[Histonet] Mouse lung IHC false positive/background issue

[Margaryan, Naira]

I have same problem for other Abs using in on mouse lung tissues... My secondary and tertiary have no problems on other tissues for negative controls as well....

Naira
**************
Confidentiality Note: This message (including any attachments) is intended for the use of the named recipient(s) only and may contain confidential and/or proprietary information. If you are not the intended recipient, please contact the sender and delete this message. Any unauthorized use of the information contained in this message is prohibited.


Ranked nationally in all 10 pediatric specialties by U.S. News & World Report  (LCHOC Ver 1.0)

This message contains confidential information and is intended only for the individual named. If you are not the named addressee you should not disseminate, distribute or copy this e-mail. Please notify the sender immediately by e-mail if you have received this e-mail by mistake and delete this e-mail from your system. E-mail transmission cannot be guaranteed to be secure or error-free as information could be intercepted, corrupted, lost, destroyed, arrive late or incomplete, or contain viruses. The sender therefore does not accept liability for any errors or omissions in the contents of this message, which arise as a result of e-mail transmission. If verification is required please request a hard-copy version.  (LCHOC VER 1.0)
-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu [mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Lu Ze
Sent: Friday, October 11, 2013 2:38 PM
To: histonet <@t> lists.utsouthwestern.edu
Cc: rebeliza <@t> gmail.com
Subject: [Histonet] Mouse lung IHC false positive/background issue

Hello everyone,

I forwarded message from my colleague, Rebecca (see below) to see if any one with IHC experience on mouse lung tissue have suggestion.


===========================================================================================

I'm having trouble with my IHC. I am staining for human MTDH (specific for human) on mouse tumor and lung sections from my xenograft mouse model. All of my tumors stained positive, but not the lung sections (which could possibly be the results). My real problem is that my negative control normal lung section stained positive for MTDH. I saw positive staining on the tissue near the outside and in the bronchioles.

I know the problem is not the secondary antibody because I did 2 slides (normal lung and previously positive tumor) without primary ab and they showed no positive staining.

My antibody is a rabbit monoclonal antibody at 1:200 for 1 hour. I'm blocking with 1% BSA in TBST for 1 hour. I'm using DAB substrate kit for developing.

What would you recommend?

Thank you,

Ze

================================
Ze Lu, Ph.D.
Optimum Therapeutics, LLC
9363 Towne Centre Dr., Suite 110
San Diego, CA 92121
_______________________________________________
Histonet mailing list
Histonet <@t> lists.utsouthwestern.edu
http://lists.utsouthwestern.edu/mailman/listinfo/histonet