Human C-Peptide Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
DICP00
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Human C-Peptide ELISA Standard Curve
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Human C-Peptide Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
2.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (10 uL)
Sensitivity
2.88 pmol/L
Assay Range
28.9 - 1,850 pmol/L (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
Specificity
Natural and synthetic human Insulin C-Peptide.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human Insulin C-Peptide Immunoassay is a 2.5 hour solid-phase ELISA designed to measure human Insulin C-Peptide in cell culture supernates, serum, plasma, and urine. It contains synthetic human Insulin C-Peptide and has been shown to accurately quantitate the peptide. Results obtained using natural human Insulin C-Peptide showed linear curves that were parallel to the standard curves obtained using the Quantikine kit standards. These results indicate that this kit can be used to determine relative mass values for naturally occurring human Insulin C-Peptide.

Precision

Intra-Assay Precision (Precision within an assay) Intra-assay Precision (Precision within an assay)<br>Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Inter-assay Precision (Precision between assays)<br>Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 129 436 845 127 437 861
Standard Deviation 5.77 14.3 13.2 12.4 33 65.3
CV% 4.5 3.3 1.6 9.8 7.6 7.6

Recovery

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
Serum (n=4) 103 96-109
Urine (n=4) 97 81-106

Linearity

To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human Insulin C-Peptide were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human C-Peptide ELISA Linearity

Data Examples

Human C-Peptide ELISA Standard Curve

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: C-Peptide

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.

Long Name:
Proinsulin C-Peptide
Entrez Gene IDs:
3630 (Human); 16333 (Mouse); 397415 (Porcine); 280829 (Bovine); 483665 (Canine)
Alternate Names:
C-P; C-Peptide; IDDM2; ILPR; Insulin C-Peptide; insulin; IRDN; MODY10; Proinsulin C-Peptide; proinsulin
⚠ WARNING: This product can expose you to chemicals including N,N-Dimethylforamide, which is known to the State of California to cause cancer. For more information, go to www.P65Warnings.ca.gov.

Assay Procedure

Refer to the product for complete assay procedure.

Bring all reagents and samples to room temperature before use. It is recommended that all samples, standards, and controls be assayed in duplicate.
  1.   Prepare all reagents, standard dilutions, and samples as directed in the product insert.
  2.   Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 1 hour.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 1 hour.
  10.   Aspirate and wash 4 times.

  11. 200 µL Substrate Solution
  12.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes. PROTECT FROM LIGHT.

  13. 50 µL Stop Solution
  14.   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.

FAQs

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ELISA Controls

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