Detection of Recombinant Mce1C and Mce1D Proteins

Western blot was carried out with our earlier protocol with slight modifications^{18 (link)}. The recombinant E. coli expressing Mce1C and Mce1D proteins were induced with 0.2 mmol/l IPTG. After sonication and centrifugation, the precipitation was transferred onto polyvinylidene fluoride membranes. Mice and rabbits were immunized subcutaneously with N. farcinica, and the antisera were used as primary antibodies and anti-rabbit IgG (Beyotime Biotechnology, China), or anti-mouse IgG antibodies (SouthernBiotech, USA) were used as secondary antibodies to detect recombinant Mce1C and Mce1D proteins.

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Ji X., Zhang X., Sun L., Hou X., Song J., Tan X., Song H., Qiu X., Li M., Tang L., Han L, & Li Z. (2020). Mce1C and Mce1D facilitate N. farcinica invasion of host cells and suppress immune responses by inhibiting innate signaling pathways. Scientific Reports, 10, 14908.

Publication 2020

anti-rabbit IgG anti-mouse IgG antibodies

Anti igg Antibodies Antisera Centrifugation E coli Iptg Membranes Mouse Polyvinylidene fluoride Proteins Rabbit Rabbits Western blot

Corresponding Organization :

Other organizations : Shandong Provincial Hospital, Shandong First Medical University, Capital Medical University, National Institute for Communicable Disease Control and Prevention, National Institute for Viral Disease Control and Prevention, Chenzhou First People's Hospital, First Affiliated Hospital of Chengdu Medical College

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Variable analysis

independent variables

Induction of recombinant E. coli expressing Mce1C and Mce1D proteins with 0.2 mmol/l IPTG

dependent variables

Detection of recombinant Mce1C and Mce1D proteins using Western blot

control variables

Sonication and centrifugation of recombinant E. coli cells
Transfer of precipitation onto polyvinylidene fluoride membranes
Use of mice and rabbits immunized subcutaneously with N. farcinica as source of primary antibodies
Use of anti-rabbit IgG and anti-mouse IgG antibodies as secondary antibodies

controls

Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned

Annotations

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