Journal
/
Biochemistry
/
Titration ELISA as a Method to Determine the Dissociation Constant of Receptor Ligand Interaction
JoVE Journal
Biochemistry
A subscription to JoVE is required to view this content.  Sign in or start your free trial.
JoVE Journal Biochemistry
Titration ELISA as a Method to Determine the Dissociation Constant of Receptor Ligand Interaction

Titration ELISA as a Method to Determine the Dissociation Constant of Receptor Ligand Interaction

DOI:
10.3791/57334-v

•

12:38 min

•

February 15, 2018

•

1Institute of Physiological Chemistry and Pathobiochemistry,University of Münster

Chapters

  • 00:04Title
  • 00:45Immobilization of the Receptor (Integrin α2A-domain) to a Microtiter Plate
  • 02:09Preparation of a Serial Dilution Row of the Ligand (Rhodocetin)
  • 03:25Binding of Ligand at Different Concentrations to Immobilized Receptor
  • 05:03Quantification of Receptor-bound Ligand by ELISA
  • 07:06Evaluation of the Titration Signals
  • 09:30Results: Rhodocetin Binds with Lower Affinity to Mutant Integrin α2A-domain
  • 10:58Conclusion

Summary

Automatic Translation

Automatic Translation

A detailed protocol to perform a titration ELISA is described. Moreover, a novel algorithm is presented to evaluate titration ELISAs and to obtain a dissociation constant of binding of a soluble ligand to a microtiter plate-immobilized receptor.

Tags

Keywords: Titration ELISADissociation ConstantReceptor-ligand InteractionIntegrin Alpha 2 A DomainRhodocetinAffinity ConstantProtein-ligand InteractionReceptor ResearchPharmaceutical ResearchPharmacology

Article

Embed

ADD TO PLAYLIST

Usage Statistics

Related Videos

check_url/57334?article_type=v

Method for Identifying Small Molecule Inhibitors of the Protein-protein Interaction Between HCN1 and TRIP8b
check_url/57334?article_type=v

Fluorescence Anisotropy as a Tool to Study Protein-protein Interactions
check_url/57334?article_type=v

Utilizing the Ethylene-releasing Compound, 2-Chloroethylphosphonic Acid, as a Tool to Study Ethylene Response in Bacteria
check_url/57334?article_type=v

A Simple Method for High Throughput Chemical Screening in Caenorhabditis Elegans
check_url/57334?article_type=v

Method for Efficient Refolding and Purification of Chemoreceptor Ligand Binding Domain
check_url/57334?article_type=v

A Colorimetric Method for Measuring Iron Content in Plants
check_url/57334?article_type=v

Extracellular Protein Microarray Technology for High Throughput Detection of Low Affinity Receptor-Ligand Interactions
check_url/57334?article_type=v

Visualizing Surface T-Cell Receptor Dynamics Four-Dimensionally Using Lattice Light-Sheet Microscopy
check_url/57334?article_type=v

A Planarian Motility Assay to Gauge the Biomodulating Properties of Natural Products
check_url/57334?article_type=v

A High-Throughput Enzyme-Coupled Activity Assay to Probe Small Molecule Interaction with the dNTPase SAMHD1

Read Article