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Hi, A customer is enquiring about the use of “EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit” with a primary antibody which is raised in rat (rat anti mouse CD144 - IgG1). He wants to know if there is any way he can use the Expose kit? I suggested that the only way is to use an extra step ie a rabbit anti rat secondary followed by the goat anti rabbit-HRP (from the expose kit). Has anyone done this before? Is there any reason why this would not work. In any case, he does not have any rabbit anti rat on hand – if that would work, can you suggest a suitable one he could buy? Thanks. Kind regards,
Asked on Dec 09 2011
Thank you for contacting us. The EXPOSE Mouse and Rabbit Specific HRP/DAB Detection IHC kit (ab80436) can be used to detect mouse and rabbit primary antibodies. You are therefore correct in thinking that the kit would need to be modified in order to detect a Rat primary antibody. This could potentially be done by using a different secondary antibody to "change the species" or the primary antibody, although we do not have any experimental data regarding this, in theory it should work. The concentration of the rabbit anti-rat antibody would need to be optimized by the user, I would recommend they try ~10 ug/ml to start. I would also suggest that the customer should purchase the ab80437 kit instead of the ab80436 kit, since they do not require the rabbit anti-mouse antibody for their application. When choosing the antibody to use your customer has to be a bit careful as the kit only detects antibodies of the IgG isotype. An antibody that could be used for this application is ab102205 which is a rabbit polyclonal (IgG) to rat IgG. I hope this information has been of help. If you require any further information please do not hesitate to ask.
Abcam Scientific Support
Answered on Dec 12 2011