Cortisol ELISA Kit

Sample Cortisol Conc. (pg/mL) %CV 1 1,174.3 6.0 2 475.9 5.6 3 177.4 14.7 Sample Cortisol Conc. (pg/mL) %CV Sample Cortisol Conc. (pg/mL) %CV 1 1,174.3 6.0 1 1,174.3 6.0 2 475.9 5.6 2 475.9 5.6 3 177.4 14.7 3 177.4 14.7 Sample Cortisol Conc. (pg/mL) %CV 1 1,188.1 7.2 2 508.7 6.3 3 199.7 10.9 Sample Cortisol Conc. (pg/mL) %CV Sample Cortisol Conc. (pg/mL) %CV 1 1,188.1 7.2 1 1,188.1 7.2 2 508.7 6.3 2 508.7 6.3 3 199.7 10.9 3 199.7 10.9

The DetectX® Cortisol Immunoassay Kit is designed to quantitatively measure cortisol present in dried fecal extracts, saliva, urine, serum, plasma and tissue culture media samples. Please read the complete kit insert before performing this assay. Total cortisol is measured in extracted samples and in serum and plasma and free cortisol in saliva and urine. A cortisol standard is provided to generate a standard curve for the assay and all samples must be read off a user-generated standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with an antibody to capture mouse antibodies. A cortisolperoxidase conjugate is added to the wells. The binding reaction is initiated by the addition of a monoclonal antibody to cortisol. The immunological reaction occurs between the limiting amount of added anti-cortisol monoclonal antibody, the cortisol antigen in the sample or standard, and the limiting amount of added cortisol-peroxidase conjugate. As the concentration of cortisol in the sample increases, the amount of cortisol-peroxidase conjugate bound decreases causing an decrease in signal, and vice versa. The signal is generated from the cortisol-peroxidase bound to the anti-cortisol antibody which itself is bound to the goat anti-mouse IgG coated plates. Excess cortisol-peroxidase does not bind to the plates and is washed out of the well prior to the addition of substrate. For a further explanation go to: www.ncbi.nlm.nih.gov/books/NBK92434/pdf/ immunometh.pdf, page 28. After an hour incubation the plate is washed and substrate is added. The substrate reacts with the bound cortisol-peroxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader at 450 nm wavelength. The concentration of the cortisol in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.

Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Reconstitution & Storage Instructions Reconstitution & Storage Instructions Western Blotting/Immunoblotting (WB/IB) Protocol Western Blotting/Immunoblotting (WB/IB) Protocol Immunohistochemistry (IHC) Protocol Immunohistochemistry (IHC) Protocol Immunocytochemistry (ICC) Protocol Immunocytochemistry (ICC) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Enzyme-Linked ImmunoSorbent Assay (ELISA) Protocol Blocking Peptide Competition Protocol (BPCP) Blocking Peptide Competition Protocol (BPCP) Immunoprecipitation (IP) Protocol Immunoprecipitation (IP) Protocol Antibody Array (AA) Protocol Antibody Array (AA) Protocol

Component Quantity Clear Coated 96 Well Plate 1 or 5 each Cortisol Standard 125 uL or 625 uL DetectX® Cortisol Antibody 3 mL or 13 mL DetectX® Cortisol Conjugate 3 mL or 13 mL Assay Buffer Concentrate 28 mL or 55 mL Dissociation Reagent 1 mL or 5 mL Wash Buffer Concentrate 30 mL or 125 mL TMB Substrate 11 mL or 55 mL Stop Solution 5 mL or 25 mL Plate Sealer 1 or 5 Component Quantity Component Quantity Clear Coated 96 Well Plate 1 or 5 each Clear Coated 96 Well Plate 1 or 5 each Cortisol Standard 125 uL or 625 uL Cortisol Standard 125 uL or 625 uL DetectX® Cortisol Antibody 3 mL or 13 mL DetectX® Cortisol Antibody 3 mL or 13 mL DetectX® Cortisol Conjugate 3 mL or 13 mL DetectX® Cortisol Conjugate 3 mL or 13 mL Assay Buffer Concentrate 28 mL or 55 mL Assay Buffer Concentrate 28 mL or 55 mL Dissociation Reagent 1 mL or 5 mL Dissociation Reagent 1 mL or 5 mL Wash Buffer Concentrate 30 mL or 125 mL Wash Buffer Concentrate 30 mL or 125 mL TMB Substrate 11 mL or 55 mL TMB Substrate 11 mL or 55 mL Stop Solution 5 mL or 25 mL Stop Solution 5 mL or 25 mL Plate Sealer 1 or 5 Plate Sealer 1 or 5