[Preparation and purification of polyclonal antibody against Toxoplasma gondii microneme protein 16 and its application in subcellular localization].

OBJECTIVE

To prepare and purify the rabbit anti-TgMIC16 polyclonal antibody, so as to apply it in subcellular localization.

METHODS

New Zealand white rabbits were immunized with purified recombinant TgMIC16 mixing with the same volume of Freund's adjuvant for three times, respectively. The rabbit serum was collected on the 14th day after the last immunization. The polyclonal antibody in rabbit serum was purified with Protein A affinity purification column. ELISA and Western blotting were used to detect the antibody titer and specificity of polyclonal antibody. The polyclonal antibody was used to the localization of TgMIC16 by the immunofluorescence method.

RESULTS

Indirect ELISA showed that the antibody titer was 1∶512 000. Western blotting showed that the recombinant TgMIC16 protein was recognized by the specific polyclonal antibody. IFA showed that TgMIC16 was located in the microneme of Toxoplasma gondii.

CONCLUSIONS

The rabbit anti-TgMIC16 is prepared and purified, and successfully applied to immunofluorescence localization of TgMIC16 in T. gondii.