Human sICAM-3 ELISA Assay


The Human sICAM-3 ELISA Assay is to be used for the in-vitro quantitative determination of soluble Inter Cellular Adhesion Molecule -3 (sICAM-3) in human serum, plasma, buffered solutions or cell culture medium. The assay will recognize both natural and recombinant human sICAM-3. The Eagle Biosciences Human CD50 / sICAM-3 ELISA Assay kit is for Research Use Only.

SKU: C5031-K01 Categories: , ,

Human sICAM-3 ELISA Assay

The Human sICAM-3 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.17 ng/ml
Dynamic Range: 0.31 ng/ml – 10 ng/ml
Incubation Time: 2h 45 min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: Human CD50 ELISA, Soluble Intercellular Adhesion Molecule 3

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.
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.
EDTA, citrate and heparin plasma can be assayed. Spin samples at 1000 x g for 30 min to remove particulates. Harvest plasma.
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

The intercellular adhesion molecule 3 (ICAM 3), also called CD50 (Cluster of Differentiation 50), is a protein which, in humans, is encoded by the ICAM 3 gene.

The protein encoded by this gene belongs to the family of intercellular adhesion molecules (ICAM). All ICAM proteins are type I transmembrane glycoproteins, contain 2 to 9 immunoglobulin type C2 domains, and bind to LFA-1 leukocyte adhesion protein. This protein is constitutively and abundantly expressed by all leukocytes and may be the most important ligand for LFA-1 in initiating the immune response. It functions not only as an adhesion molecule, but also as a powerful signaling molecule.

Related Products

Human CD54 ELISA Assay
Human CD102 sICAM-2 ELISA Assay
Human CD106 / sVCAM-1 ELISA Assay Kit

Additional Information

Assay Principle

The sICAM-3 Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for ICAM-3 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known sICAM-3 concentrations, control specimens and unknowns are pipetted into these wells. During the first incubation, the sICAM-3 antigen and a biotinylated monoclonal antibody specific for ICAM-3 are simultaneously incubated.

After washing, the enzyme ( streptavidin-peroxydase ) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution which is acting on the bound enzyme is added to induce a coloured reaction product. The intensity of this coloured product is directly proportional to the concentration of sICAM-3 present in the samples.

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

Typical Standard Curve

Human CD50 / sICAM-3 ELISA Assay Kit

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