Expression,purification and polyclonal antibody preparation of human sperm protein FSCB
AIM:To express the recombinant protein and prepare the polyclonal antibody(PAb)of human sperm protein FSCB.METHODS:Two recombinant bacterium B121 stains expressing human full length FSCB and truncated FSCB(FSCBt)proteins were successfully constructed,and the FSCBt protein was purified by Ni~(2+) affinity chromatography and molecular sieve chromatography.The specific PAb of FSCB was prepared by immunizing female rabbits.RESULTS:The recombinant protein FSCB and FSCBt could be expressed stably by the recombinant bacterium and be recognized by the PAb,which relative molecular mass were about 230 000 and 30 000.The purity of purified recombinant protein FSCBt was about 95%.The titers of PAb were 1:10~5.CONCLUSION:Expression of the recombinant protein FSCB and preparation of its polyclonal antibody laid a foundation for further research of its functions in human sperm.
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万方数据
维普
