Objective To clone rat NGB gene and prepare its polyclonal antibody in order to investigate the function of neuroglobin (NGB).   Methods The total RNA was extracted from Wistar rat brain and the full length cDNA encoding NGB was obtained by RT-PCR. Confirmed by sequencing analysis

it was inserted in the prokaryotic expression vector pET-28a(+). pET-28a(+)-NGB was expressed in E.coli BL21 after its sequence and reading frame were confirmed by two restriction endonucleases and sequencing. The protein of pET-28a(+)-NGB was used to immunize New Zealand rabbits for preparing polyclonal antibody after identification and purification. The antibody titer was tested by ELISA method.   Results The titer of polyclonal antibody was 100 000. Immunoblot analysis showed the polyclonal antibody can recognize fusion protein expressed in 293 cells.   Conclusion The polyclonal antibody of NGB is prepared and can be used in further study.