Human CD124 ELISA Assay


The Human CD124 ELISA Assay is to be used for the in-vitro quantitative determination of soluble IL-4 receptor molecule (CD124)  in human serum, plasma, buffered solutions or cell culture medium. The assay will recognize both natural and recombinant human IL-4R. The Eagle Biosciences Human IL-4R ELISA Assay kit is for Research Use Only.

SKU: C2431-K01 Categories: , ,

Human CD124 / IL4-R ELISA Assay Kit

The Human CD124 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 11.5 pg/ml
Dynamic Range: 31.25 pg/ml – 1000 pg/ml
Incubation Time: 1h 45min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: IL4-R, Interleukin 4 Receptor

Specimen collection, processing & storage
Cell culture supernatants, serum, plasma or other biological samples will be suitable for use in the assay. Remove serum from the clot or red cells, respectively, as soon as possible after clotting and separation.
Cell culture supernatants: Remove particulates and aggregates by spinning at approximately 1000 x g for 10 min.
Serum: Avoid any unintentional stimulation of the cells by the procedure. Use pyrogen/endotoxin free collecting tubes. Serum should be removed rapidly and carefully from the red cells after clotting. For that, after clotting, centrifuge at approximately 1000 x g for 10 min and remove serum.
Plasma: EDTA, citrate and heparin plasma can be assayed. Spin samples at 1000 x g for 30 min to remove particulates. Harvest plasma.
Storage: If not analyzed shortly after collection, samples should be aliquoted (250-500μl) to avoid repeated freeze-thaw cycles and stored frozen at –70°C. Avoid multiple freeze-thaw cycles of frozen specimens.

Assay Background

Interleukin 4 or IL-4 is a cytokine whose role is to induce the differentiation of naive helper T cells (Th0 lymphocytes) into Th2 lymphocytes. After being activated by IL-4, Th2 cells themselves start producing IL-4. Their receptor is called the interleukin 4 receptor. The cell that initially produces IL-4 for the differentiation of Th0 has not yet been clearly identified, although recent studies suggest that basophilic polynuclear cells are the effector cells. Interleukin 4 is very close to interleukin 13 which has similar effects.

Related Products

Human CD213-alpha 2 / IL-13R alpha 2 ELISA Assay
Human sCD25 / IL-2R ELISA Assay Kit
Human CD116 ELISA Assay

Additional Information

Assay Principle

The IL-4R Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for IL-4R has been coated onto the wells of the microtiter strips provided. Samples, including standards of known IL-4R concentrations and unknowns are pipetted into these wells.  During the first incubation, the IL-4R antigen and a biotinylated monoclonal antibody specific for IL-4R are simultaneously incubated.  After washing, the enzyme ( streptavidin-peroxydase ) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of IL-4R present in the samples.

  1. Prepare Standard curve as shown in section 8.4
  2. Add 100µl of each standard, sample, control and zero (standard diluent) in duplicate to appropriate number of wells
  3. Add 50µl of diluted biotinylated anti-IL-4R to all wells
  4. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hours
  5. Remove the cover and wash the plate
  6. Add 100μl of Streptavidin-HRP solution into all wells
  7. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  8. Repeat wash step 5.
  9. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  10. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  11. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve

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