Human CD14 ELISA Assay

$560.00

The Eagle Biosciences Human CD14 ELISA Assay kit is to be used for the in-vitro quantitative determination of CD14 in serum, plasma, or cell lysates. The assay will recognize both natural and recombinant CD14.  The Eagle Biosciences Human CD14 ELISA Assay kit is for Research Use Only.

SKU: C1431-K01 Categories: , ,

Human CD14 ELISA Assay

The Human CD14 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 6 ng/ml
Dynamic Range: 6.87 – 220 ng/ml
Incubation Time: Overnight
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl


Assay Background

CD14 is described as the marker molecule for monocytes and macrophages and as an important component of the innate immune system. It exists in two forms : mCD14 a 55 kDa protein anchored to the membrane by a glycosylphosphoinositol tail (GPI) and two soluble serum proteins (sCD14) with or without the GPT anchor. The mCD14 is critical for lipopolysaccharide (LPS) dependent signal transduction, and sCD14 confers LPS sensitivity to cells lacking mCD14. CD14 is strongly expressed on cells of the myelomonocytic lineage such as monocytes and osteoclast progenitors and at a lower level on macrophages, Langerhans cells, granulocytes and microglia.

Increased sCD14 levels are associated with inflammatory infectious diseases or allergy.

CD14 is involved in clearance of gram-negative bacteria via its high affinity binding to LPS-LPB complexes. CD14 is a useful marker for enumeration of monocytes or osteoclast progenitors in peripheral blood and for monocytic and myelomonocytic acute myeloid leukaemia.


Related Products

Human CD86 ELISA Assay
Human CD21 ELISA Assay
Human CD31 ELISA Assay

Additional Information

Assay Principle


The Human CD14 ELISA Assay kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA).  A monoclonal antibody specific for CD14 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known CD14 concentrations and unknowns are pipetted into these wells.  During the first incubation, the CD14 antigen, standards and zeros are bound this is followed by a separate incubation where the biotinylated monoclonal antibody specific for CD14 is added.

After washing, the enzyme (streptavidin-peroxidase) is added. After incubation and washing to remove all the unbound enzyme, a substrate solution is added to induce a colored reaction product. The intensity of this coloured product is directly proportional to the concentration of CD14 present in the samples.

  1. Add 100µl of each standard, sample and zero (Standard Diluent) in duplicate to appropriate number of wells
  2. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 2 hours
  3. Remove the cover and wash the plate
  4. Add 50µl of diluted biotinylated anti-CD14 to all wells
  5. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 1 hour
  6. Repeat wash
  7. Add 100μl of Streptavidin-HRP solution into all wells
  8. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 30 min
  9. Repeat wash
  10. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  11. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  12. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


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