Human IL-2 HS ELISA Assay

$650.00

The Human IL-2 HS ELISA Assay is to be used for the in-vitro quantitative determination interleukin-2 (IL-2) in human sera, plasmas, buffered solutions or cell culture media. The assay will recognize both natural and recombinant human IL-2. The Eagle Biosciences Human High Sensitivity IL-2 ELISA Assay kit is for Research Use Only

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Human IL-2 HS ELISA Assay

The Human IL-2 HS ELISA Assay is For Research Use Only

Size: 1×96 wells
Sensitivity: 0.97 pg/ml
Dynamic Range: 1.87 – 60 pg/ml
Incubation Time: 2 hours
Sample Type: Serum, Plasma, Cell Culture
Sample Size: 100 µl
Alternative Name: Human IL-2 High Sensitive ELISA


Assay Background

IL-2 is a powerful immunoregulatory lymphokine produced by T-Cells in response to antigenic or mitogenic activation. IL-2 stimulates growth and differentiation of B-Cells, most T-cells, NK cells, monocytes and macrophages.

Mature IL-2 is a 15.4kDa globular glycoprotein containing 133 amino acid residues including one intrachain disulfide bond between residues 58 and 105.

Apart from its most important role to mediate antigen-specific T-lymphocyte proliferation, IL-2 modulates the expression of interferon-g and major histocompatibility antigens.

Alterations in the ability of T-cell to synthesize IL-2 have been observed in physiologic and pathologic states. Currently, IL-2 is used to enhance the immune system of patients for the treatment of cancer and infectious disease.


Storage Instructions
Store the Human IL-2 ELISA Assay kit reagents between 2 and 8°C. Immediately after use remaining reagents should be returned to cold storage (2-8°C). Expiry of the kit and reagents is stated on box front labels. The expiry of the Human IL-2 ELISA Assay kit components can only be guaranteed if the components are stored properly, and if, in case of repeated use of one component, the reagent is not contaminated by the first handling.


Related Products

Human sCD25 / IL-2R ELISA Assay Kit
Human IL-6 High Sensitivity ELISA Assay Kit
Human IL-4 ELISA Assay Kit

Additional Information

Assay Principle


The IL-2 Kit is a solid phase sandwich Enzyme Linked-Immuno- Sorbent Assay (ELISA). A monoclonal antibody specific for IL-2 has been coated onto the wells of the microtiter strips provided. Samples, including standards of known IL-2 concentrations and unknowns are pipetted into these wells.  During the first incubation, the IL-2 antigen and a biotinylated monoclonal antibody specific for IL-2 are simultaneously incubated.

After washing, the enzyme (streptavidin-peroxydase) is added. All the unbound enzyme is removed by washing and the first amplification step is performed by adding the Biotine-Tyramine reagent. Under the action of HRP, a biotine polymerisation reaction occurs in the region of the HRP linked to the detection antibody. After washing the second amplification step is performed and the polymerised biotine is revealed by a new streptavidin-HRP step. Finally after washing, the substrate is added. The intensity of this coloured product is directly proportional to the concentration of IL-2 present in the samples.

Assay Procedure


  1. Add 100μl of each standard, sample and zero (appropriate Standard Diluent Buffer) in duplicate to appropriate number of wells
  2. Add 50μl of diluted biotinylated anti-IL-2 to all wells
  3. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) with slow shaking for 1 hour
  4. Remove the cover and wash the plate
  5. Add 100μl of Streptavidin-HRP solution 1 into all wells
  6. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) with slow shaking for 20 min
  7. Repeat wash step 4.
  8. Add 100μl of diluted Amplifier to all wells
  9. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) with slow shaking for 15 min
  10. Repeat wash step 4.
  11. Add 100μl of Streptavidin-HRP solution 2 into all wells
  12. Cover with a plastic plate cover and incubate at room temperature (18 to 25°C) with slow shaking for 20 min
  13. Repeat wash step 4.
  14. Add 100μl of ready-to-use TMB Substrate Solution into all wells
  15. Incubate in the dark for 10-20 minutes* at room temperature. Avoid direct exposure to light by wrapping the plate in aluminium foil
  16. Add 100μl of H2SO4:Stop Reagent into all wells

Typical Standard Curve


Human IL-2 HS ELISA Assay

Specificity


Ten specificities were tested with concentrations higher than IL-2 curve concentrations in the Human High Sensitivity IL-2 ELISA Assay Kit.  No cross reaction was observed for concentrations ranging from 300 to 18.75 pg/ml for IL-1α and β, IL-8, IL-10, TNF-α, GrB, FasL, TRAIL, CD116 and Gp-130.

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