A testing method of nucleic acid binding protein based on biochip, comprises the following steps: 1. puts a plurality of groups solution including nucleic acid captured probes into biological sample including a plurality of nucleic acid binding protein to be test, and thus forming nucleic acid captured probe-nucleic acid binding protein complexes; such nucleic acid captured probe includes at least a segment of binding sequence which can bind with aimed nucleic acid binding protein; 2. separates such nucleic acid captured probe-nucleic acid binding protein complexes, then recoveries nucleic acid captured probes; 3. hybridizes the nucleic acid captured probes according to step 2 with a plurality of single strand blotting probes on biochip substrate; the sequence of such blotting probe compensates with such nucleic acid captured probe or one of its strand; 4. detects the result of hybridization.