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[Preparation of anti-Red antisera and its subcellular localization]
Xiao-nan Zhang1, Su-min Chen, Er-ping Luo
1Department of Biochemistry and Molecular Biology, Fourth Military Medical University, Xi'an 710032, China.
Aim:
To prepare rabbit anti-Red antisera.
Methods:
The bet, exo and gam genes of lambda phage were amplified by PCR from genomic DNA and cloned into the expression vector pDH2, respectively. Red proteins were induced to express at 42 degrees C. The expressed proteins were analyzed by PAGE and thin-layer scanning. The antisera were prepared by immunizing rabbits with the three Red proteins, respectively. The titers and specificities of the antisera were detected by Western blot.
Results:
Beta, Exo and Gam proteins accounted for about 40.3%, 49.2% and 73.4% of total bacterial protein, respectively. The titers of the antisera were about 1:2,000. Western blot analysis indicated that the three antisera all had good specificities to the corresponding proteins.
Conclusion:
Specific anti-Red antisera are prepared successfully.
