Human CD86 ELISA Assay


The Eagle Biosciences Human CD86 ELISA is to be used for the in-vitro quantitative determination of CD86 in human serum, plasma, buffered solutions or cell culture medium. The assay will recognize both natural and recombinant human CD86. The Eagle Biosciences Human CD86 ELISA Assay kit is for Research Use Only.

SKU: C8631-K01 Categories: , ,

Human CD86 ELISA Assay

The Human CD86 ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.6 U/ml
Dynamic Range: 1.187 U/ml – 38 U/ml
Incubation Time: 2h 40min
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: B7-2

Scientific Background

CD86 (also known as B7-2) is a transmembrane glycoprotein member of the IgG superfamily. CD86 is expressed on antigen-presenting cells that provide co-stimulatory signals necessary for T cell activation and survival. It is the ligand for two different proteins on the T cell surface: CD28 (for autoregulation and intercellular association) and CTLA-4 (for attenuation of regulation and cellular disassociation). CD86 works in tandem with CD80 (B7-1) to prime T cells and its co-stimulatory pathway plays an important role in the development of anti-tumour response.

Some studies have shown that CD86 expression by malignant cells is associated with a poor prognosis in both acute myeloid leukemia and multiple myeloma and a decrease of CD86 on PBM in melanoma results in an ineffective immune response.

A high level of soluble CD86 in plasma was described in several diseases: abdominal aortic aneurysm, leukaemia, systemic lupus erythematosus and acute asthma.

Related Products

Human CD95 / FAS ELISA Assay Kit
Human CD56/NCAM-1 ELISA Kit
Human CD10/Neprilysin ELISA Kit

Additional Information

Assay Principle

The CD86 Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for CD86 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known CD86 concentrations and unknowns are pipetted into these wells. During the first incubation, the CD86 antigen and a biotinylated monoclonal antibody specific for CD86 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 CD86 present in the samples.

  1. Add 100µl of each standard, sample and zero (Standard Diluent) in duplicate to appropriate number of wells
  2. Add 50µl of diluted biotinylated anti-CD86 to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) for 2 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 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

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