This human IL-2 ELISA is a 4.5 hour solid phase immunoassay readily applicable to measure IL-2 levels in serum, plasma, cell culture supernatant, and other biological fluids in the range of 0 to 2500pg/ml. It showed no cross reactivity with other cytokines tested. This IL-2 ELISA is expected to be effectively used for further investigations into the relationship between IL-2 and the various conditions mentioned.
This assay employs the quantitative sandwich enzyme immunoassay technique. A monoclonal antibody specific for IL-2 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-2 present is bound by the immobilized antibody. After washing away any unbound substances, an enzyme-linked polyclonal antibody-enzyme specific for IL-2 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-2 bound in the initial step. The color development is stopped and the intensity of the color is measured.
In order to measure the concentration of IL-2 in the samples, this kit includes two calibration diluents (Calibrator Diluent I for serum/plasma testing and Calibrator Diluent II for cell culture supernatant testing.) According to the testing system, the provided standard is diluted (2-fold) with the appropriate Calibrator Diluent and assayed at the same time as the samples. This allows the operator to produce a standard curve of Optical Density (O.D) versus IL-2 concentration (pg/ml). The concentration of IL-2 in the samples is then determined by comparing the O.D. of the samples to the standard curve.
Sensitivity: (same/more than)5.7pg/ml (using Calibrator Diluent I)
(same/more than)7.1pg/ml (using Calibrator Diluent II)
Range: 39-2500pg/ml
Kit Components:
I7663-28E1: Microtiter Plate,1x96 wells. Precoated with anti-human IL-2 Mab
*I7663-28E4: IL-2 Standard (5ng/vial), 2x1vial. Supplied as a lyophilized powder containing recombinant human Il-2 in a buffered protein base with preservative.
I7663-28E5: Calibrator Diluent I, 1x25ml. For serum/plasma testing. Supplied as animal serum with buffer and preservative.
I7663-28E6: Calibrator Diluent II, 1x25ml. For cell culture supernatant testing. Supplied as cell culture medium with calf serum and preservative.
I7663-28E7: Wash Buffer (20X), 1x60ml. Supplied as a concentrated solution of buffered surfactant.
I7663-28E8: Substrate A, 1x10ml. Supplied as a buffered solution with H2O2.
I7663-28E9: Substrate B, 1x10ml. Supplied as a buffered solution wih TMB.
I7663-28E10; Stop Solution, 1x15ml. Supplied as 2N Sulphuric Acid (H2SO4).
I7663-28E11: IL-2 Pab (Biotin)1x7ml
I7663-28E12: Avidin (HRP) 1x14ml
Storage and Stability:
Store at 4 degrees C. Stable for 6 months. *Reconstituted I7663-28E4 (stock solution) is stable for 1 month at -20 degrees C. For maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap.
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