Fluorokine Receptor Detection Kits

Fluorokine Receptor Detection Kits employ recombinant biotinylated ligands to quantitatively determine cytokine receptor density using flow cytometry. These simple kits allow the user to estimate the percentage of cells within a population which express the receptor of interest. In addition, cell surface receptor density can be assessed.

Cells are incubated with the biotinylated cytokine corresponding to the receptor of interest. Binding of the recombinant biotinylated ligand to cell surface receptors directly mirrors the physiological ligand/receptor interaction. Cell samples are then incubated with fluorescein-conjugated avidin. Cells expressing the specific cytokine receptors are fluorescently stained, with the intensity of staining proportional to the density of the receptors. Relative receptor density is determined by flow cytometric analysis using 488 nm wavelength laser excitation.

Fluorokine Receptor Detection Kits include reagents to support the specificity of the results. All kits contain a control protein (i.e. soybean trypsin inhibitor) biotinylated to the same degree as the ligand of interest. In addition, species specific antibodies (supplied in the kit) can be employed to inhibit ligand/receptor interaction, demonstrating the protein specificity of the binding of the biotinylated cytokine. Non-conjugated ligands can also be applied to provide further support for reaction specificity.

Features

  • Determine the percentage of cells bearing receptors within a population
  • Estimate the receptor density on cell surfaces
  • Can be used with peripheral blood cells and cultured cells
  • Biotinylated ligands mirror physiological ligand/receptor interactions
  • Negative control provided in the form of biotinylated control proteins
  • Pretreatment with blocking antibody supports the protein-specificity of the binding
  • Binding specificity can be further supported by applying non-conjugated ligand (not included)

Kit Contents

  • Biotinylated recombinant, species specific ligand
  • Avidin conjugated with fluorescein (at an f:p ratio of 5:1)
  • Negative control reagents
  • Blocking antibody
  • Wash buffer concentrate

Assay Principle

Kits to determine receptor density using flow cytometry.
Assay Pinciple

Step 1

Cell samples are incubated with the biotinylated recombinant ligand.

Step 2

Fluorescein-conjugated avidin is applied and complexes with biotinylated ligand.

Step 3

Unbound materials are washed away. Relative receptor density is determined by flow cytometry using 488 nm wavelength laser excitation.

Control experiments to confirm specificity

All kits contain a control protein (i.e. soybean trypsin inhibitor) biotinylated to the same degree as the ligand of interest. In addition, species specific antibodies (supplied in the kit) can be employed to inhibit ligand/receptor interaction, demonstrating the protein specificity of the binding of the biotinylated cytokine. Non-conjugated ligands can also be applied to provide further support for reaction specificity.