The invention discloses a method for separating
copper proteome by using
copper-chelated magnetic beads, and relates to the technical field of
protein chemistry. The method comprises the following steps: taking a
biological tissue or
cell, after fully
grinding with
liquid nitrogen, adding
protein extracting liquid, and after uniformly mixing, carrying out low-temperature
centrifugation, so as to obtain a denatured
protein solution; mixing the
copper-chelated magnetic beads with the denatured
protein solution, and incubating to obtain a
magnetic bead protein
mixed solution; adding a
buffer solution 1 with the volume 1 time of that of the
magnetic bead protein
mixed solution into the
magnetic bead protein
mixed solution for suspension, pouring half of the supernate under the external
magnetic field, and repeating for 4-5 times to obtain a mixed solution A; adding a
buffer solution 2 with the volume 1 time of that of the mixed solution A into the mixed solution A, separating all the specific copper-binding proteins, namely, the copper
proteome, and collecting the copper
proteome under the external
magnetic field. According to the method, the metalloproteome is separated under a denaturing condition by using magnetic bead-IDA, so that the procedures are greatly simplified, and the obtaining efficiency of metalloproteins is improved; a method suitable for separating the metalloproteome from the tissue or
cell under the excess heavy
metal treatment condition is established.