CGMP ELISA Kit

Sample Cyclic GMP Conc. (pmol/mL) %CV 1 16.0 5.7 2 10.9 4.7 3 7.6 5.5 Sample Cyclic GMP Conc. (pmol/mL) %CV Sample Cyclic GMP Conc. (pmol/mL) %CV 1 16.0 5.7 1 16.0 5.7 2 10.9 4.7 2 10.9 4.7 3 7.6 5.5 3 7.6 5.5 Sample Cyclic GMP Conc. (pmol/mL) %CV 1 15.4 7.0 2 10.2 6.8 3 7.0 6.5 Sample Cyclic GMP Conc. (pmol/mL) %CV Sample Cyclic GMP Conc. (pmol/mL) %CV 1 15.4 7.0 1 15.4 7.0 2 10.2 6.8 2 10.2 6.8 3 7.0 6.5 3 7.0 6.5 Sample Cyclic GMP Conc. (pmol/mL) %CV 1 10.0 5.5 2 7.1 3.9 3 1.5 7.1 Sample Cyclic GMP Conc. (pmol/mL) %CV Sample Cyclic GMP Conc. (pmol/mL) %CV 1 10.0 5.5 1 10.0 5.5 2 7.1 3.9 2 7.1 3.9 3 1.5 7.1 3 1.5 7.1 Sample Cyclic GMP Conc. (pmol/mL) %CV 1 9.7 15.8 2 6.3 10.2 3 3.8 15.4 Sample Cyclic GMP Conc. (pmol/mL) %CV Sample Cyclic GMP Conc. (pmol/mL) %CV 1 9.7 15.8 1 9.7 15.8 2 6.3 10.2 2 6.3 10.2 3 3.8 15.4 3 3.8 15.4

The DetectX® Direct Cyclic GMP (cGMP) Immunoassay kit is designed to quantitatively measure cGMP present in lysed cells, EDTA plasma, urine, saliva and tissue culture media samples. Please read the complete kit insert before performing this assay. For samples where the levels of cGMP are expected to be relatively high, the regular format for the assay can be used. For samples with expected low levels of cGMP, an optional acetylation protocol can be used. The kit is unique in that all samples and standards are diluted into an acidic Sample Diluent, which contains special additives and stabilizers, for cGMP measurement. This allows plasma, urine and saliva samples to be read in an identical manner to lysed cells. Acidified samples of cGMP are stable and endogenous phosphodiesterases are inactivated in the Sample Diluent. A cGMP standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. A clear microtiter plate coated with an antibody to capture rabbit IgG is provided and a neutralizing Plate Primer solution is added to all the used wells. Standards or diluted samples, either with or without acetylation, are pipetted into the primed wells. A cGMP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a rabbit polyclonal antibody to cGMP to each well. After a 2 hour incubation, the plate is washed and substrate is added. The substrate reacts with the bound cGMPperoxidase conjugate. After a short incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. The concentration of the cGMP 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 Coated Clear 96 Well Plates 1 or 5 Each Cyclic GMP Standard 125 uL or 625 uL DetectX® Cyclic GMP Antibody 3 mL or 13 mL DetectX® Cyclic GMP Conjugate 3 mL or 13 mL Sample Diluent Concentrate 12 mL or 60 mL Plate Primer 25 mL Acetic Anhydride 2 mL Triethylamine 4 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 Each Component Quantity Component Quantity Coated Clear 96 Well Plates 1 or 5 Each Coated Clear 96 Well Plates 1 or 5 Each Cyclic GMP Standard 125 uL or 625 uL Cyclic GMP Standard 125 uL or 625 uL DetectX® Cyclic GMP Antibody 3 mL or 13 mL DetectX® Cyclic GMP Antibody 3 mL or 13 mL DetectX® Cyclic GMP Conjugate 3 mL or 13 mL DetectX® Cyclic GMP Conjugate 3 mL or 13 mL Sample Diluent Concentrate 12 mL or 60 mL Sample Diluent Concentrate 12 mL or 60 mL Plate Primer 25 mL Plate Primer 25 mL Acetic Anhydride 2 mL Acetic Anhydride 2 mL Triethylamine 4 mL Triethylamine 4 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 Each Plate Sealer 1 or 5 Each