GUCA2A ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Guanylate Cyclase Activator 2A (GUCA2A) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Guanylate Cyclase Activator 2A (GUCA2A) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

GCAP-I; Guanylate cyclase activator 2A; guanylate cyclase activator 2A (guanylin 2, intestinal, heat-stable) ; guanylate cyclase-activating protein 1; guanylate cyclase-activating protein I; guanylin; GUCA2; prepro-guanylin; STARA.

Endogenous activator of intestinal guanylate cyclase. It stimulates this enzyme through the same receptor binding region as the heat-stable enterotoxins.

The test principle applied in this kit is Sandwich enzyme immunoassay. The microtiter plate provided in this kit has been pre-coated with an antibody specific to Guanylate Cyclase Activator 2A (GUCA2A) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Guanylate Cyclase Activator 2A (GUCA2A) . Next, Avidin conjugated to Horseradish Peroxidase (HRP) is added to each microplate well and incubated. After TMB substrate solution is added, only those wells that contain Guanylate Cyclase Activator 2A (GUCA2A), biotin-conjugated antibody and enzyme-conjugated Avidin will exhibit a change in color. The enzyme-substrate reaction is terminated by the addition of sulphuric acid solution and the color change is measured spectrophotometrically at a wavelength of 450nm +/- 10nm. The concentration of Guanylate Cyclase Activator 2A (GUCA2A) in the samples is then determined by comparing the O.D. of the samples to the standard curve.

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

Matrix Recovery Range (%) Average (%) Serum (n=5) 94-101 97 EDTA Plasma (n=5) 81-97 94 Heparin Plasma (n=5) 78-103 98 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) Serum (n=5) 94-101 97 Serum (n=5) 94-101 97 EDTA Plasma (n=5) 81-97 94 EDTA Plasma (n=5) 81-97 94 Heparin Plasma (n=5) 78-103 98 Heparin Plasma (n=5) 78-103 98

Component Amount Anti-GUCA2A Microplate 96 Wells (12 x 8 Well Strips) GUCA2A Lyophilized Standard 2 x 1 ng 100X Biotinylated GUCA2A Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Component Amount Component Amount Anti-GUCA2A Microplate 96 Wells (12 x 8 Well Strips) Anti-GUCA2A Microplate 96 Wells (12 x 8 Well Strips) GUCA2A Lyophilized Standard 2 x 1 ng GUCA2A Lyophilized Standard 2 x 1 ng 100X Biotinylated GUCA2A Detector Antibody 120 uL 100X Biotinylated GUCA2A Detector Antibody 120 uL Avidin/HRP Conjugate 120 uL Avidin/HRP Conjugate 120 uL Standard Diluent 1 x 20 mL Standard Diluent 1 x 20 mL Detector Antibody Diluent 1 x 12 mL Detector Antibody Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL Conjugate Diluent 1 x 12 mL 30X Wash Buffer 1 x 20 mL 30X Wash Buffer 1 x 20 mL TMB Substrate 1 x 9 mL TMB Substrate 1 x 9 mL Stop Solution 1 x 6 mL Stop Solution 1 x 6 mL

This assay has high sensitivity and excellent specificity for detection of Guanylate Cyclase Activator 2A (GUCA2A) . No significant cross-reactivity or interference between Guanylate Cyclase Activator 2A (GUCA2A) and analogues was observed.