I sometimes have to use multiple different antibodies that are unfortunately produced in the same species (like rabbit)...I would say it can work so long as you strip your blot appropriately, thus you could likely have success using multiple rabbit antibodies such that you don't get signal from the previous antibodies, its also a plus if the molecular weight targets are different too.

Alternately, if applicable for your experiments, you could cut your blot into strips and probe each strip with the different rabbit antibodies so that you do not need to re-probe using multiple antibodies of the same species.

Thank you doctor@ Aaron Dhanda you mean it is useful if your target have different M.WT or I can cut the blot to avoid cross reactivity of using multiple antirabbit 2ndry Abs.

Faheem Hezam Al-Mughales it is useful yes but not a requirement, the usefulness is that if they have different molecular weights then you can be quite certain you are not getting signal from the other antibodies that you have tested since recall, even if you strip your blot, there still may be some molecular amounts of antibody left on the blot.

If you are just testing different rabbit antibodies with your lysate and do not need to run specific controls for each lane in your gel then yes I would say load your lysate into your gel, and if its is for instance a 10 lane gel, then after you transfer the gel onto your blot, simply cut the blot into 2 or 3 or 4 strips..doing so now you can easily probe each strip with a different rabbit antibody and so do not need to run the potentially minor risk of stripping and re-probing with multiple rabbit antibodies.