Primary cultured urothelial cells (UT) isolated from normal and FIC cat bladders (n=3 per group) were assessed for evidence of endogenous release of stress-related peptides CRF and Ucn. The plated cells were allowed to stabilize and grow for two days in vitro (2DIV) prior to commencement. The culture medium was carefully removed from the culture wells, replaced with sterile filtered HBSS and the dishes replaced in the incubator. At 30 minutes the medium was removed to investigate the presence of CRF and urocortin. Enzyme Immunoassay (EIA) kits from Phoenix Pharmaceuticals (Burlingame, CA; EK-019-06 and EK-019-15, detect CRF and non-selectively Ucn I, II and III, respectively) following the manufacturer’s instructions. The linear detection range of both kits was 0.1–4ng/ml. CRF Immunoassay has been validated against prepro-CRF (125–151), PACAP-38, LH-RH, ACTH, [Arg8] Vasopressin, and BNP45. UCN Immunoassay is 100% cross-reactive with UCN I, II, and III and has been validated against cortistatin-14, CRF, MCH, LH-RH, NPY, and somatostatin. Prior to assay, peptides were extracted from the cell culture supernatant using C-18 SEP Columns (Phoenix Pharmaceuticals, Burlingame, USA) following the manufacturer’s instructions.
Enzyme immunoassay eia kits
Manufactured by Phoenix Pharmaceuticals
Sourced in United States
Enzyme Immunoassay (EIA) kits are a type of laboratory equipment used for the detection and quantification of various analytes, such as proteins, hormones, and other biomolecules, in biological samples. EIA kits employ the principles of immunoassay, where a specific antibody is used to capture and detect the target analyte. The assay involves the use of enzymes that catalyze a color-producing reaction, allowing for the quantification of the analyte through colorimetric or fluorometric measurements.
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Lab products found in correlation
2 protocols using enzyme immunoassay eia kits
1
Urothelial Cells Stress Peptide Release
2
Glucose, Insulin, GLP-1, and GIP Dynamics
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The levels of plasma glucose, serum insulin, plasma total GLP-1, and plasma total GIP were determined for each time point. Besides, we calculated the incremental area under the curves of plasma glucose (iAUC-glucose), serum insulin (iAUC-insulin), plasma total GLP-1 (iAUC-GLP-1), and plasma total GIP (iAUC-GIP) using the values from 0 to 180 min based on the trapezoidal rule. Serum insulin was measured by chemiluminescent enzyme immunoassay (SRL, Tokyo, Japan). Plasma total GLP-1 and GIP were measured in duplicate using the enzyme immunoassay (EIA) kits (Phoenix Pharmaceuticals, Burlingame, CA, USA), and intra- and inter-assay coefficients of variation in the EIA were < 10 and < 15%, respectively. HbA1c, total cholesterol, and triglyceride were measured by enzymatic assay (SRL, Tokyo, Japan), and HDL cholesterol was measured directly (SRL, Tokyo, Japan).
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