Application notes:
EliKine™ Rat IL-4 ELISA Kit employs a two-site sandwich ELISA to quantitate IL-4 in samples. An antibody specific for IL-4 has been pre-coated onto a microplate. Standards and samples are pipetted into the wells and any IL-4 present is bound by the immobilized antibody. After removing any unbound substances, a biotin-conjugated antibody specific for IL-4 is added to the wells. After washing, proprietary EliKine™ Streptavidin-HRP conjugates is added to the wells. Following a wash to remove any unbound streptavidin-enzyme reagent, a substrate solution is added to the wells and color develops in proportion to the amount of IL-4 bound in the initial step. The color development is stopped and the intensity of the color is measured.
IL-4 rat ELISA Kit components:
1. Rat IL-4 microplate: 96 well polystyrene microplates (12 strips of 8wells) coated with the antibody specific for Rat IL-4.
2. Rat IL-4 standard: Rat IL-4 in a buffered protein base with preservatives, lyophilized.
3. Standard diluent: Diluent solution for dilute standards and samples.
4. Assay Buffer: Diluent solution for dilute biotin-conjugated antibody and Streptavidin-HRP.
5. Biotin-Conjugate Rat IL-4 detection antibody: Biotin-conjugated Rat IL-4 detection antibody. 100× liquid.
6. EliKine™ Streptavidin-HRP: 100× liquid.
7. HRP substrate: TMB (Tetramethyl-benzidine) solution.
8. Stop solution: 2N sulfuric acid.
9. Wash buffer: PBS with 1%Tween-20; 20× liquid.
10. Plate covers.
Assay protocol summary:
Typical data:
Detection range: 1.56 pg/mL-100 pg/mL
Sensitivity: The minimum detectable dose (MDD) of Rat IL-4 is typically less than 1.0 pg/mL
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