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Related Experiment Videos

A rapid, high capacity assay for basic fibroblast growth factor binding

W F Herblin1, F A Barbera

  • 1DuPont Merck Pharmaceutical Company, Wilmington, DE 19880-0400, USA.

Life Sciences
|March 3, 1995
PubMed
Summary
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A new assay for basic fibroblast growth factor (bFGF) binding was developed using rat lung membranes. This high-capacity assay is useful for screening compounds and identifying inhibitors of bFGF binding.

Area of Science:

  • Biochemistry
  • Molecular Biology

Background:

  • Basic fibroblast growth factor (bFGF) plays crucial roles in cell growth and tissue repair.
  • Developing efficient assays for bFGF binding is essential for drug discovery and understanding its biological functions.

Purpose of the Study:

  • To develop a rapid, high-capacity assay for measuring basic fibroblast growth factor (bFGF) binding.
  • To validate the assay's utility in screening compounds and characterizing inhibitors.

Main Methods:

  • Utilized rat lung tissue membranes and 2M sodium chloride to remove endogenous bFGF.
  • Adapted the assay to a 96-well Millipore Multi-Screen system for high-throughput screening.
  • Validated the assay using known inhibitors like suramin and protamine sulfate.

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Main Results:

  • The assay demonstrated high capacity and efficiency in a 96-well format.
  • Standard inhibitors (suramin, protamine sulfate) validated the assay's performance.
  • Dimercaptothiadiazole showed increased potency against high-affinity bFGF binding.

Conclusions:

  • The developed assay is a valuable tool for screening potential inhibitors of bFGF.
  • The assay allows for the differentiation between high-affinity and overall binding inhibition.
  • This method aids in the detailed examination of suspected bFGF inhibitors.