Human CD54 ELISA Assay


The Human sICAM-1/CD54 ELISA is to be used for the in-vitro quantitative determination of soluble Inter Cellular Adhesion Molecule -1 (sICAM-1) in human serum, plasma, buffered solutions or cell culture medium. The assay will recognize both natural and recombinant human sICAM-1/CD54. The Human sICAM-1/CD54 ELISA Assay kit is for Research Use Only.

SKU: C5431-K01 Categories: , ,

Human CD54 ELISA Assay

The Human CD54 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.1 ng/ml
Dynamic Range: 0.25 ng/ml – 8 ng/ml
Incubation Time: 1h 40min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: ICAM-1 ELISA, Intercellular Adhesion Molecule 1 ELISA

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 clothing. For that, after clothing, 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

ICAM-1 (InterCellular Adhesion Molecule) or CD54 is a protein encoded by the human ICAM1 gene located on human chromosome 19. It is present on the surface of endothelial cells and lymphocytes. It can serve as a receptor for viruses of the genus rhinovirus.

The protein encoded by this gene is a type of continuous intercellular adhesion molecule present at low concentrations in leukocyte membranes and endothelial cells. During cytokine stimulation, the concentrations increase considerably. When activated, leukocytes bind to endothelial cells via ICAM-1 / LFA-1 and then transmigrate into tissues.

Related Products

Human CD44 ELISA Assay Kit
Human CD31 ELISA Assay Kit
Human CD21 ELISA Assay Kit

Additional Information

Assay Principle

The CD54 Kit is a solid phase sandwich Enzyme Linked-Immuno-Sorbent Assay (ELISA). A monoclonal antibody specific for CD54 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known CD54 concentrations and unknowns are pipetted into these wells. During the first incubation, the CD54 antigen and a biotinylated monoclonal antibody specific for CD54 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 CD54 present in the samples.

Assay Procedure

  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-CD54 to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hours
  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 5.
  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

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