A colorimetric method for detection of specific ligand binding
| Autor: | Suzanne K. Beckner, Hubert Wang |
|---|---|
| Rok vydání: | 1992 |
| Předmět: |
Streptavidin
Phosphatase Biophysics Biotin Ligands Biochemistry Cell Line chemistry.chemical_compound Bacterial Proteins Cell surface receptor Animals Humans Receptor Molecular Biology Chemistry Ligand binding assay Cell Membrane Receptors Interleukin-2 Cell Biology Ligand (biochemistry) Alkaline Phosphatase Molecular biology Phosphoric Monoester Hydrolases Receptors Interleukin-3 Kinetics Biotinylation Alkaline phosphatase Interleukin-2 Colorimetry Interleukin-3 |
| Zdroj: | Analytical biochemistry. 204(1) |
| ISSN: | 0003-2697 |
| Popis: | The binding of IL-2 and IL-3 to the factor-dependent cell lines CTB6 and 32D, respectively, was determined using biotinylated ligand detected by the addition of a streptavidin/alkaline phosphatase conjugate and amplified with a phosphatase amplification system. Binding of both ligands was detectable after incubation with as little as 20 fmol of ligand and could be inhibited with a 10-fold molar excess of nonbiotinylated ligand. No binding was observed when biotinylated ligand was incubated with a receptor negative cell line (PC-12) and IL-2 was unable to compete with biotinylated IL-3 binding to 32D cells, further demonstrating specificity. These studies indicate that biotinylated ligands can be used as a nonradioactive method to detect specific, high-affinity cell surface receptors. |
| Databáze: | OpenAIRE |
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