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Related Experiment Videos

Sample centrifugation onto membranes for sequencing.

D Sheer1

  • 1Applied Biosystems Inc., Foster City, California 94404.

Analytical Biochemistry
|May 15, 1990
PubMed
Summary
This summary is machine-generated.

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This study introduces a novel centrifugation method for adsorbing proteins onto polyvinylidene difluoride (PVDF) membranes. The technique efficiently concentrates and purifies proteins, even in low picomole quantities, for subsequent analysis.

Area of Science:

  • Biochemistry
  • Analytical Chemistry
  • Proteomics

Background:

  • Protein adsorption onto membranes is crucial for various biochemical assays.
  • Existing methods may face limitations in efficiency and sample recovery.

Purpose of the Study:

  • To develop and validate a new centrifugation-based method for protein adsorption onto PVDF membranes.
  • To enhance protein concentration, purification, and recovery for downstream analyses.

Main Methods:

  • Utilized a dual-membrane system (PVDF and low molecular weight cut-off) within a microcentrifuge tube.
  • Employed centrifugation to drive protein adsorption onto the PVDF membrane.
  • Validated the method with varying protein sample sizes (micrograms to picomoles) and analyzed recovery via radioactivity and amino acid analysis.

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Main Results:

  • Demonstrated efficient protein recovery across a range of sample quantities.
  • Showcased effective removal of interfering substances like buffers and sodium dodecyl sulfate.
  • Achieved useful sequence information from low picomole samples after purification.

Conclusions:

  • The developed centrifugation method offers an efficient and versatile approach for protein adsorption and purification.
  • This technique is suitable for handling small protein quantities and preparing samples for sensitive analyses like protein sequencing.