Protein-specific immunodetection of A unphosphorylated HSP-60 protein, and B phosphorylated p44 and p42 mitogen-activated protein kinase (MAPK) proteins. F: flow-through; E: eluate fractions. The antibody used to detect MAPK recognizes an epitope containing phosphorylated residues at Thr202 and Tyr204 in the p44 (upper band) and p42 (lower band) MAPK amino acid sequences. The absence of unphosphorylated HSP-60 in the eluate fraction and the absence of phosphorylated MAPK in the flow-through fraction demonstrate the complete separation of phosphorylated proteins using the PhosphoProtein Purification Kit.
