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Author Topic: non-specific preserum staining  (Read 2941 times)

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Offline zhuliangru

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non-specific preserum staining
« on: February 13, 2006, 12:43:43 PM »
:?
Hi! I am doing IHC with paraffin sections of human trachea. The first antibody is peptide-specific rabbit antiserum. I got strong staining for trachea epithelium. However, non-specific preimmune serum also have staining, although needed more time to get the same intensity.

My questions is how to slove this non-specific staining issue with negative control serum, will further dilution of antibody help? Also, do you think if I use IgG fraction instead of the serum will help to solve the non-specific staining?

Thanks a lot!

non-specific preserum staining
« on: February 13, 2006, 12:43:43 PM »

Offline richard03

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non-specific preserum staining
« Reply #1 on: February 16, 2006, 11:22:51 PM »
Make sure your primary ab concentration is same as the preimmune serum. Try to dilute the ab more will help to reduce background as well.

Richard

Offline ImmunoNYC

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Re: non-specific preserum staining
« Reply #2 on: February 19, 2006, 12:54:46 PM »
This is a well known area of endogenous biotin background. Are you using biotin protocol by chance???? What I am saying is that your problem may not lie with your antibody and preserum?

With more details of your protocol I am sure we could help.


Quote from: "zhuliangru"
I got strong staining for trachea epithelium. However, non-specific preimmune serum also have staining, although needed more time to get the same intensity.

Re: non-specific preserum staining
« Reply #2 on: February 19, 2006, 12:54:46 PM »