Domain-Specific Monoclonal Antibodies Against Human Rev-erbβ

Area of Science:

• Molecular Biology
• Immunology
• Gene Regulation

Background:

• The nuclear receptor Rev-erbβ is a critical transcriptional factor.
• Functional studies of Rev-erbβ have been hindered by the absence of specific antibodies.

Purpose of the Study:

• To generate and characterize monoclonal antibodies (MAbs) against human Rev-erbβ.
• To develop tools for investigating the biological roles of Rev-erbβ.

Main Methods:

• Immunization of mice with purified recombinant human Rev-erbβ-6His fusion protein.
• Generation of hybridoma cell lines secreting anti-Rev-erbβ MAbs.
• Western blotting, immunofluorescence, and immunoprecipitation assays to assess antibody reactivity and specificity.

Main Results:

• Five hybridoma cell lines producing anti-Rev-erbβ MAbs were successfully generated.
• All five MAbs demonstrated strong immunoreactivity against recombinant Rev-erbβ.
• One MAb, 37H8, exhibited the ability to immunoprecipitate endogenous Rev-erbβ.
• Antibody cross-reactivity with Rev-erbα was observed for antibodies specific to shared C and E domains.

Conclusions:

• The developed MAbs are valuable tools for studying Rev-erbβ.
• The MAb 37H8 is particularly useful for detecting endogenous Rev-erbβ.
• These antibodies will facilitate further research into Rev-erbβ's functions in biological systems.