Human C-Peptide Quantikine ELISA Kit Summary
Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine
|Intra-Assay Precision||Inter-Assay Precision|
The recovery of human Insulin C-Peptide spiked to levels throughout the range of the assay in various matrices was evaluated.
|Sample Type||Average % Recovery||Range %|
|Cell Culture Media (n=4)||99||84-106|
|EDTA Plasma (n=4)||101||95-108|
|Heparin Plasma (n=4)||102||90-115|
Human C-Peptide ELISA Standard Curve
Preparation and Storage
Insulin and the IGFs are members of the insulin family of molecules. IGF-I and II are structurally homologous to proinsulin, and can be thought of as taking the shape of an exaggerated, inverted letter G. Intrachain disulfide bonds link sections of the G. The single chain insulin propeptide consists of a 30 amino acid B chain (aa 2554), a C-peptide (aa 5589), and a 21 aa A chain (aa 90110). Removal of the C-peptide by proteolysis enables the formation of mature Insulin, a disulfidelinked heterodimer of the A and B chains. Circulating C-peptide levels are elevated in hyperinsulinism, obesity, and type II diabetes.
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1 hour.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.
- Add 50 µL of Stop Solution to each well. Read at 450 nm within 30 minutes. Set wavelength correction to 540 nm or 570 nm.
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Appropriate to detect human c-peptide in rabbit samples. Tested to contaminate the standard curve with rabbit serum and anterior chamber fluid of the eye with neglectable interference as compared to vehicle (diluent provided by the kit).