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The nuclear autoimmune antigen Ku is also present on the cell surface
R G Dalziel1, S C Mendelson, J P Quinn
1Department of Veterinary Pathology, Royal (Dick) School of Veterinary Studies, Summerhall, Edinburgh.
Autoimmunity
|January 1, 1992
Summary
Polyclonal antibodies were developed against the 85 and 70 kDa subunits of the Ku complex. These antibodies successfully identified Ku proteins in both whole cell extracts and on the cell membrane.
Area of Science:
- Immunology
- Molecular Biology
- Cell Biology
Background:
- The Ku complex is a crucial protein heterodimer involved in DNA repair.
- Understanding the specific subunits and their cellular localization is important for comprehending DNA repair mechanisms.
Purpose of the Study:
- To generate and characterize polyclonal antibodies against the 85 and 70 kDa subunits of the Ku complex.
- To validate the specificity and utility of these antibodies in identifying Ku proteins within cellular compartments.
Main Methods:
- Purification of 85 and 70 kDa Ku complex subunits from MLA144 T cell nuclear extract.
- Western blot analysis using generated antibodies on HeLa cell whole cell extracts.
- Flow cytometry (FACS) analysis to detect Ku proteins on the cell membrane.
Main Results:
- Successfully raised polyclonal antibodies against the individual 85 and 70 kDa Ku subunits.
- Antibodies specifically recognized the target Ku subunits in Western blot analysis of HeLa cell extracts.
- Antibodies demonstrated the ability to identify Ku proteins located on the cell membrane via FACS analysis.
Conclusions:
- The generated polyclonal antibodies are specific and effective tools for detecting Ku complex subunits.
- These antibodies can be utilized to study Ku protein expression and localization in various cellular contexts, including nuclear and membrane fractions.