Currently, I work with a kinase which itself also gets phosphorylated. It needs to get phosphorylated for activation. Probing with the phospho-kinase antibody for a specific residue like Threonine gives multiple bands around 2 or 3 in the molecular weight marker range. Can anybody guide me concerning it if they had done this experiment, which among the protein bands would be correct? Also, I probed with the total kinase after stripping the blots. After merging both images using Image Lab software, I could get one band among the phospho-kinase covering the region of total kinase. Is that band among the phospho-kinase the original band for that targeted region or the band above?