SCNN1A ELISA Kit (Human)

Intra-assay Precision (Precision within an assay) : 3 samples with low, middle and high level Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) were tested 20 times on one plate, respectively. Inter-assay Precision (Precision between assays) : 3 samples with low, middle and high level Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) were tested on 3 different plates, 8 replicates in each plate. CV (%) = SD/meanX100 Intra-Assay: CV<10% Inter-Assay: CV<12%

Alpha ENaC-2; alpha-ENaC; alpha-NaCH; amiloride-sensitive epithelial sodium channel alpha subunit; amiloride-sensitive sodium channel subunit alpha; amiloride-sensitive sodium channel subunit alpha 2; BESC2; ENaCa; ENaCalpha; epithelial Na (+) channel subunit alpha; LIDLS3; nasal epithelial sodium channel alpha subunit; nonvoltage-gated sodium channel 1 subunit alpha; SCNEA; SCNN1; sodium channel epithelial 1 alpha subunit; sodium channel, non voltage gated 1 alpha subunit; sodium channel, nonvoltage-gated 1 alpha; sodium channel, non-voltage-gated 1 alpha.

Sodium permeable non-voltage-sensitive ion channel inhibited by the diuretic amiloride. Mediates the electrodiffusion of the luminal sodium (and water, which follows osmotically) through the apical membrane of epithelial cells. Plays an essential role in electrolyte and blood pressure homeostasis, but also in airway surface liquid homeostasis, which is important for proper clearance of mucus. Controls the reabsorption of sodium in kidney, colon, lung and eccrine sweat glands. Also plays a role in taste perception.

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 Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) . Standards or samples are then added to the appropriate microtiter plate wells with a biotin-conjugated antibody specific to Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) . 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 Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a), 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 Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) 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) 80-89 83 EDTA plasma (n=5) 88-96 92 heparin plasma (n=5) 81-103 95 Matrix Recovery Range (%) Average (%) Matrix Recovery Range (%) Average (%) serum (n=5) 80-89 83 serum (n=5) 80-89 83 EDTA plasma (n=5) 88-96 92 EDTA plasma (n=5) 88-96 92 heparin plasma (n=5) 81-103 95 heparin plasma (n=5) 81-103 95

Component Amount Anti-SCNN1A Microplate 96 Wells (12 x 8 Well Strips) SCNN1A Lyophilized Standard 2 x 10 ng 100X Biotinylated SCNN1A 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-SCNN1A Microplate 96 Wells (12 x 8 Well Strips) Anti-SCNN1A Microplate 96 Wells (12 x 8 Well Strips) SCNN1A Lyophilized Standard 2 x 10 ng SCNN1A Lyophilized Standard 2 x 10 ng 100X Biotinylated SCNN1A Detector Antibody 120 uL 100X Biotinylated SCNN1A 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 Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) . No significant cross-reactivity or interference between Amiloride Sensitive Sodium Channel Subunit Alpha (SCNN1a) and analogues was observed.