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Preparative electrophoresis in a concentrated polymer solution: automated procedure for microsome isolation
H T Chang1, N Chen, J V Sullivan
1Department of Chemistry, National Taiwan University, Taipei, Republic of China.
Analytical Biochemistry
|April 5, 1997
Summary
Automated gel electrophoresis now isolates large particles. This modified HPGE 1000 apparatus restores computer-directed recovery for particles too big for gels, enabling quantitative isolation.
Area of Science:
- Biotechnology
- Analytical Chemistry
- Biophysics
Background:
- Automated gel electrophoresis (HPGE 1000) offers preparative capabilities with fluorescence-monitored electroelution.
- Previous applications of this preparative technique were limited to subcellular particles in polymer solutions.
- Manual positioning of the recovery chamber hindered full automation in prior methods.
Purpose of the Study:
- To detail modifications for restoring automated operation to the HPGE 1000 apparatus.
- To extend the benefits of automated quantitative recovery to larger particles.
- To enable isolation of particles that cannot enter standard electrophoresis gels.
Main Methods:
- Modification of the HPGE 1000 apparatus instrumentation and procedure.
- Development of a method for computer-directed positioning of the recovery chamber.
- Application of stepwise extended electroelution time with fluorescence monitoring.
Main Results:
- Successful restoration of automated operation for particle isolation.
- Demonstration of quantitative recovery for particles too large to enter gels.
- Validation of the modified procedure for enhanced preparative electrophoresis.
Conclusions:
- The modified HPGE 1000 apparatus successfully automates the isolation of large particles.
- This advancement expands the utility of automated electroelution beyond gel electrophoresis.
- The procedure offers a quantitative and automated method for isolating challenging particle sizes.