C-Peptide ELISA Kit (Rat)

ELISA Kit

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C-PEPTIDE

C-peptide is a protein that is produced in the body along with insulin. First preproinsulin is secreted with an A-chain, C-peptide, a B-chain, and a signal sequence. The signal sequence is cut off, leaving proinsulin. Then the C-peptide is cut off, leaving the A-chain and B-chain to form insulin. Cellular effects of C-peptide: C-peptide has been shown to bind to the surface of a number of cell types such as neuronal, endothelial, fibroblast and renal tubular, at nanomolar concentrations to a receptor that is likely G-protein-coupled. The signal activates Ca2+-dependent intracellular signaling pathways such as MAPK, PLCγ, and PKC, leading to upregulation of a range of transcription factors as well as eNOS and Na+K+ATPase activities.

Assay Methodology: Quantitative Competitive ELISA

Aviva Systems Biology C-Peptide ELISA Kit (Rat) (OKEH03700) is based on a competitive enzyme immuno assay technique. The microtiter well-plate in this kit has been pre-coated with an anti-human C-Peptide antibody. Sample or standards are added to the wells along with a fixed quantity of biotinylated C-Peptide and incubated. The C-Peptide found in the sample or standards competes with the biotinylated C-Peptide for limited binding sites on the immobilized anti-human C-Peptide antibody. Excess unbound biotinylated C-Peptide and sample or standard C-Peptide is washed from the plate. Avidin-HRP conjugate is added, incubated and washed. An enzymatic reaction is then produced through the addition of TMB substrate which is catalyzed by the immobilized HRP to generate a blue color product that changes yellow after adding acidic stop solution. The density of yellow coloration is measured by reading the absorbance at 450 nm which is quantitatively proportional to the amount of biotinylated C-Peptide captured in the well and inversely proportional to the amount of C-Peptide which was contained in the sample or standard.

Serum, Plasma, Tissue Homogenates, Cell Culture Supernates, Other Biological Fluids