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Development of a fluorescent ligand-binding assay using the AcroWell filter plate.

Publication ,  Journal Article
Valenzano, KJ; Miller, W; Kravitz, JN; Samama, P; Fitzpatrick, D; Seeley, K
Published in: J Biomol Screen
December 2000

One of the most powerful tools for receptor research and drug discovery is the use of receptor-ligand affinity screening of combinatorial libraries. Early work involved the use of radioactive ligands to identify a binding event; however, there are numerous limitations involved in the use of radioactivity for high throughput screening. These limitations have led to the creation of highly sensitive, nonradioactive alternatives to investigate receptor-ligand interactions. Pall Gelman Laboratory has introduced the AcroWell, a patented low-fluorescent-background membrane and sealing process together with a filter plate design that is compatible with robotic systems. Taken together, these allow the AcroWell 96-well filter plate to detect trace quantities of lanthanide-labeled ligands for cell-, bead-, or membrane-based assays using time-resolved fluorescence. Using europium-labeled galanin, we have demonstrated that saturation binding experiments can be performed with low-background fluorescence and signal-to-noise ratios that rival traditional radioisotopic techniques while maintaining biological integrity of the receptor-ligand interaction. In addition, the ability to discriminate between active and inactive compounds in a mock galanin screen is demonstrated with low well-to-well variability, allowing reliable determination of positive hits even for low-affinity interactions.

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Published In

J Biomol Screen

DOI

ISSN

1087-0571

Publication Date

December 2000

Volume

5

Issue

6

Start / End Page

455 / 461

Location

United States

Related Subject Headings

  • Receptors, Neuropeptide
  • Receptors, Galanin
  • Radioligand Assay
  • Medicinal & Biomolecular Chemistry
  • Ligands
  • Iodine Radioisotopes
  • Humans
  • Galanin
  • Fluorescent Dyes
  • Fluorescence
 

Citation

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Valenzano, K. J., Miller, W., Kravitz, J. N., Samama, P., Fitzpatrick, D., & Seeley, K. (2000). Development of a fluorescent ligand-binding assay using the AcroWell filter plate. J Biomol Screen, 5(6), 455–461. https://doi.org/10.1177/108705710000500608
Valenzano, K. J., W. Miller, J. N. Kravitz, P. Samama, D. Fitzpatrick, and K. Seeley. “Development of a fluorescent ligand-binding assay using the AcroWell filter plate.J Biomol Screen 5, no. 6 (December 2000): 455–61. https://doi.org/10.1177/108705710000500608.
Valenzano KJ, Miller W, Kravitz JN, Samama P, Fitzpatrick D, Seeley K. Development of a fluorescent ligand-binding assay using the AcroWell filter plate. J Biomol Screen. 2000 Dec;5(6):455–61.
Valenzano, K. J., et al. “Development of a fluorescent ligand-binding assay using the AcroWell filter plate.J Biomol Screen, vol. 5, no. 6, Dec. 2000, pp. 455–61. Pubmed, doi:10.1177/108705710000500608.
Valenzano KJ, Miller W, Kravitz JN, Samama P, Fitzpatrick D, Seeley K. Development of a fluorescent ligand-binding assay using the AcroWell filter plate. J Biomol Screen. 2000 Dec;5(6):455–461.
Journal cover image

Published In

J Biomol Screen

DOI

ISSN

1087-0571

Publication Date

December 2000

Volume

5

Issue

6

Start / End Page

455 / 461

Location

United States

Related Subject Headings

  • Receptors, Neuropeptide
  • Receptors, Galanin
  • Radioligand Assay
  • Medicinal & Biomolecular Chemistry
  • Ligands
  • Iodine Radioisotopes
  • Humans
  • Galanin
  • Fluorescent Dyes
  • Fluorescence