检测鸡TRIM25蛋白的免疫组化方法

doi:10.11843/j.issn.0366-6964.2018.04.027

Abstract

Abstract:

The experiment was conducted to set up the immunohistochemitry assay to detect chicken TRIM25 expression in the chicken's tissues. Chicken TRIM25 gene was amplified from chicken's spleen and the gene was cloned into a prokaryotic expression vector pEASY-Blunt E1 to construct recombinant plasmid named as pEASY-Blunt E1-TRIM25. The recombinant plasmid was expressed in E. coli and then identified by SDS-PAGE and Western blot. After being purified, the fusion protein was inoculated to rabbits and BALB/c mice as antigen for preparing polyclonal antibody. The titer of the antibody was detected by ELISA, and the sera antibodies were used to setup immunohistochemistry assay; the tissue sections of chicken's lung and spleen and liver were assayed. The results showed that TRIM25 was expressed as fusion protein with 48 kD. The titers of the antibody in rabbits' and mice's sera were above 1:10⁶. The results of immunohistochemistry assay indicated that TRIM25 mainly exists in the cytoplasm of lymphocytes, hepatocyte, and epithelial cells in chicken's lung, spleen and liver. In conclusion, the immunohistochemitry assay mediated with polyclonal antibody against chicken TRIM25 were successfully set up in this study and TRIM25 distribution in some tissues of chickens was firstly reported.

CLC Number:

S852

CHU Ying, MA Wen-jie, HU Wei-guo, CHEN Ru, QIU Jian-hua, GUO Hui-jun, LI Hong-mei. Immunohistochemistry Assay for Detecting Chicken TRIM25 in Different Tissues[J]. ACTA VETERINARIA ET ZOOTECHNICA SINICA, 2018, 49(4): 865-870.

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Immunohistochemistry Assay for Detecting Chicken TRIM25 in Different Tissues

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