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The Protein Block in the kit does not contain any rabbit serum. The laboratory hasnot tested this kit with the addition of a rabbit serum blocking step. However,in theory if the source of the background is non-specific binding of the goat anti-rabbit HRP conjugate of the kit, than blocking with rabbit serum would possibly increase the background. Indeed,the goat anti-rabbit HRP would bind to any rabbit antibody bound to the tissue (including the primary). We would therefore recommend to use a goat serum for blocking to test whether it will reduce the background.
Please let me know if this helps. I would be happy to help to troubleshoot the protocol to improve the staining. Please do send then an image and the detailled protocol as well as the mention of the controls used. Thank you. I look forward to hear back from you.
Abcam Scientific Support
Answered on Aug 24 2012