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In Vitro Assays And Screening List

In Vitro Assays And Screening List

Specializing in both standard and custom in vitro assays, we are focused on providing the right assays for target identification & validation, assay development, screening, hit-to-lead validation to help identify candidate compounds for preclinical and clinical development.

Primary & Secondary Radioligand Binding Assays

Compounds are typically subjected to primary radioligand binding assays at targets (receptors) selected by investigators. In the primary binding assays, compounds are usually tested at single concentrations (10 μM) in quadruplicate in 96-well plates. Compounds that show a minimum of 50% inhibition at 10 μM are tagged for secondary radioligand binding assays to determine equilibrium binding affinity at specific targets.

Both primary and secondary radioligand binding assays are carried out in a final of volume of 125 μl per well in appropriate binding buffer. The hot ligand concentration is usually at a concentration close to the Kd (unless stated otherwise). Total and nonspecific binding are determined in the absence and presence of 10 μM of the appropriate reference compound, respectively. In brief, plates are incubated at a specific temperature in the dark for a specific amount of time (receptor specific).

Reactions are stopped by vacuum filtration onto 0.3% polyethyleneimine (PEI) soaked 96-well Unifilters using a Filtermate harvester, followed by a wash with cold wash buffer. Scintillation cocktail is then added onto air-dried filters. Radioactivity is counted in a Microbeta counter.


Key details of our functional binding assay services:

  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Gold standard filtration assays.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Over 100 targets covered.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Single concentration screening or dose-response for IC50 determinations.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Percent inhibition (mean of replicates).
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Individual values as a percent of control, IC50 value (calculated from a minimum of 6 concentration testing).
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Plotted IC50 curves.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Proven skills and expertise to develop difficult targets.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Reliable, quick turnaround times.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 High quality data with strict quality controls.
  • Created by potrace 1.10, written by Peter Selinger 2001-2011 Flexible custom services to supplement our off-the-shelf services to meet your needs, providing a cost-effective way to expand your capabilities and capacity.

How We Work

Brainstorming

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Research

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Report

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Case Studies

Compound Characterization

Compound Characterization

Compound Characterization Study Goal Determine ex vivo receptor occupancy of CMPDx at Orexin-1 (OX1) receptors using radiolabeled Orexin antagonist Created by potrace 1.10, written by Peter Selinger 2

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Compound Characterization 1

Compound Characterization 1

Compound Characterization Study Goal Determine ex vivo receptor occupancy of CMPDx at Orexin-1 (OX1) receptors using radiolabeled Orexin antagonist Created by potrace 1.10, written by Peter Selinger 2

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