Ang 1 enzyme immunoassay eia kit

Control and AGT KO mice were placed in metabolic cages for 18 consecutive days for measurement of food and water intake, body weight and 24‐h urine collection. Mice were given 9 mL of a normal (4 days), high (7 days), or low Na⁺ (7 days) gelled diet with free access to water. On days 4, 11, and 18, 35 μL of blood was collected for assay of plasma renin concentration and plasma AGT. Plasma renin concentration (PRC) was measured as the amount of angiotensin‐1 (Ang‐I) generated after incubation with excess porcine angiotensinogen using the Ang‐I enzyme immunoassay (EIA) kit (Bachem, San Carlos, CA). Plasma and urine total AGT were measured using a commercially available EIA kit (IBL America, Minneapolis, MN). Urinary Na⁺ and K⁺ were determined using the EasyVet Analyzer (Medica, Bedford, MA) and albumin excretion using ELISA kit (Albuwell M kit, Exocell Inc). At the conclusion of metabolic balance studies, mice were sacrificed and kidneys isolated for further analyses.

PRC measurement was performed on the 7th day of normal salt (NS), low salt (LS), or high salt (HS) diets. For hemorrhagic shock experiments, PRC was measured at 0, 1, 3, 9, 15 and 20 hours post hemorrhagic shock as previously reported [16 (link)]. The mice were lightly anesthetized with 1.5% isoflurane. Blood samples (~25 μL/sample) were collected from the retro-orbital plexus and centrifuged for 10 minutes at 1500 × g. Plasma samples (~10 μL) were obtained and incubated with excess porcine angiotensinogen (0.4 μmol/L; Sigma, St. Louis, MO, USA) for 20 min. The amount of angiotensin (Ang) I generated after incubation was used to calculate the PRC with the Ang I enzyme immunoassay (EIA) kit (Bachem, San Carlos, CA, USA). PRC was expressed as the amount of Ang I generated per hour per milliliter of plasma.