[Histonet] Mouse lung IHC false positive/background issue

[Lu Ze]

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. 


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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

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Ze Lu, Ph.D.
Optimum Therapeutics, LLC
9363 Towne Centre Dr., Suite 110
San Diego, CA 92121