Related Experiment Video
Updated: Aug 8, 2026
Generation of Murine Monoclonal Antibodies by Hybridoma Technology
Published on: January 2, 2017
Generation and characterization of a monoclonal antibody, mH1, raised against mitotic HeLa cells
M A Okubo1, S Chiba, T Nishikata
1Department of Biological Science, Faculty of Science, Hiroshima University, Higashi-Hiroshima, Japan.
Insights
Researchers developed a monoclonal antibody (mH1) that targets specific structures on dividing HeLa cells. This antibody identifies a gamma-actin homolog, offering new insights into cell division mechanisms.
Area of Science:
- Cell Biology
- Immunology
- Molecular Biology
Background:
- Hybridoma technology enables the production of specific monoclonal antibodies.
- Understanding cell division requires identifying proteins involved in specific cellular structures.
Purpose of the Study:
- To generate a monoclonal antibody targeting mitotic HeLa cells.
- To characterize the localization and targets of the obtained antibody.
Main Methods:
- Hybridoma cell line preparation and immunization.
- Immunofluorescence and immunoelectron microscopy for antigen localization.
- Immunoblot analysis and N-terminal amino acid sequencing for protein identification.
Main Results:
- A monoclonal antibody (mH1) was generated, reacting strongly with cleavage furrows of dividing HeLa cells.
- mH1 antigens were localized to microvillus projections on dividing HeLa cells.
- mH1 recognized 42-kDa and 130-kDa protein components; the 42-kDa component is a gamma-actin homolog.
Conclusions:
- Monoclonal antibody mH1 is a valuable tool for studying cell division.
- The identified gamma-actin homolog plays a role in the surface structures of dividing cells.
Abstract:
Hybridoma cell lines were prepared from spleen cells of mouse immunized with mitotic HeLa cells. A monoclonal antibody (mH1), which intensively reacted with cleavage furrows of dividing HeLa cells in immunofluorescence, was obtained. In interphase, this antibody diffusely stained whole HeLa cells. Immunoelectron microscopy showed that mH1 antigens were localized at microvillus projections at the surface of dividing HeLa cells, but definite localization of that antigen was not observed in interphasic cells. Immunoblot analysis showed that mH1 is reactive to 42-kDa and 130-kDa components. Further, the 42-kDa component was identified as a gamma-actin homolog by N-terminal amino acid sequence analysis.
