The ELISA method is a benchmark for quantitation of pathological antigens and there are indeed many variations to this method. ELISAs are adaptable to high-throughput screening because results are rapid, consistent and relatively easy to analyze. The best results have been obtained with the sandwich format, utilizing highly purified, prematched capture and detector antibodies. The resulting signal provides data which is very sensitive and highly specific.
This section is intended to represent the main material contained in Sinobiological Inc.ELISA kit.
1.Microplate-96 well polystyrene microplate (12 strips of 8 wells) coated with the antibody against target protein.
2.Detecion Antibody-detecion antibody conjugated to horseradish peroxidase (HRP) with preservatives.
3.Standard-standard in a buffer with preservatives, lyophilized.
4.Wash Buffer Concentrate-25 mL of a 20-fold concentrated solution of buffered surfactant with preservatives.
5.Dilution Buffer Concentrate-8 mL of a 20-fold concentrated dilution buffer with preservatives.
6.Color Reagent A-13 mL of stabilized hydrogen peroxide.
7.Color Reagent B-13 mL of stabilized chromogen (tetramethylbenzidine).
8.Stop Solution-8 mL of 2 N sulfuric acid.
1.Microplate reader capable of measuring absorbance at 450 nm
2.Pipettes and pipette tips.
3.Deionized or distilled water.
4.Multi-channel pipette, squirt bottle, manifold dispenser, or automated microplate washer.
5.500 mL graduated cylinder.
6.Tubes for standard dilution.
7.Well plate cover or seals.