Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

An automated Western blot analysis using the phastsystem.

B Prieur1, F Russo-Marie

  • 1Unité Associée Institut Pasteur/INSERM No. 285, Institut Pasteur, Paris, France.

Analytical Biochemistry
|August 1, 1988
PubMed
Summary

This study introduces a fast, automated Western blot technique for analyzing minimal protein samples. The method uses the Phastsystem for efficient protein separation, transfer, and antibody detection, reducing total preparation time.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Patient perspectives following initiation of elexacaftor-tezacaftor-ivacaftor in people with cystic fibrosis and advanced lung disease.

Respiratory medicine and research·2021
Same author

Isolation by distance and sharp discontinuities in gene frequencies: implications for the phylogeography of an alpine insect species, Carabus solieri.

Molecular ecology·2004
Same author

Cytokine modulation of liver annexin 1 expression during experimental endotoxemia.

The American journal of pathology·2001
Same author

Mitochondrial membrane potential and apoptosis peripheral blood monocytes in severe human sepsis.

American journal of respiratory and critical care medicine·2001
Same author

Annexin A5 D226K structure and dynamics: identification of a molecular switch for the large-scale conformational change of domain III.

FEBS letters·2001
Same author

CREB is involved in mouse annexin A1 regulation by cAMP and glucocorticoids.

European journal of biochemistry·2000

Area of Science:

  • Biochemistry
  • Molecular Biology
  • Proteomics

Background:

  • Western blotting is a crucial technique for protein analysis.
  • Traditional Western blotting can be time-consuming and labor-intensive.
  • Analyzing small protein quantities presents technical challenges.

Purpose of the Study:

  • To develop a rapid and automated method for Western blot preparation.
  • To adapt the Phastsystem for efficient protein separation and transfer.
  • To enable the analysis of very small protein amounts.

Main Methods:

  • Utilized a commercially available electrophoresis system (Phastsystem, Pharmacia).
  • Adapted the system for gradient-sodium dodecyl sulfate gel electrophoresis and transfer to nitrocellulose.

Related Experiment Videos

  • Automated antibody incubation, blocking, and washing steps using the Phastsystem's developing unit.

    • Main Results:

    • Achieved rapid protein separation, transfer, and staining.
    • Total preparation time reduced to approximately 6–7 hours.
    • Successfully prepared Western blots from very small protein samples.

    Conclusions:

    • The developed automated method significantly accelerates Western blot preparation.
    • The Phastsystem is effective for automated, high-throughput protein analysis.
    • This technique is suitable for researchers working with limited sample quantities.