Human C-Peptide EIA

RayBio® Human C-Peptide EIA Kit optimized for serum and cell culture medium. Competition-based ELISA on a 96-well strip plate.


Antigen Information

Accession Number:
  • P01308
Gene ID:
  • 3630
Gene Symbols:
  • INS
Protein Name & Synonyms:
Insulin [Cleaved into: Insulin B chain Insulin A chain]

Assay Format

Detection Range:
0.1-1,000 ng/ml
Recommended Dilution (serum & plasma):
Human: 2X
772 pg/ml
This kit only detects C-peptide, not Insulin A or B chain.

Cross Reactivity: This EIA kit shows no cross-reactivity with any of the cytokines tested: Ghrelin, Nesfatin, Angiotensin II, NPY and APC.
Number of Targets Detected:
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Serum
Design Principle:
  • Competition-based
Method of Detection:
  • Quantitative
Solid Support:
96-well Microplate

Product Specifications

1, 2, or 5 x 96-Well Strip Microplate Kit


C-Peptide is a 31-amino acid single-chain peptide that is made when proinsulin is split into insulin and C-Peptide. They split before proinsulin is released from endocytic vesicles within the pancreas-one C-Peptide for each insulin molecule. C-Peptide is the middle segment of proinsulin that is between the N-terminal B-chain (30 amino acids) and the C-terminal A-chain (21 amino acids).

Unlike insulin, C-Peptide has no known physiological function. Due to the fact that C-Peptide has a 2-to-5-time longer half-life than insulin, there are higher concentrations of C-Peptide than insulin in the peripheral circulation. Therefore, plasma C-Peptide concentrations may reflect pancreatic insulin secretion more reliably than the level of insulin itself. C-Peptide is cleared from the body by the kidney, and, unlike insulin, urine concentrations are 20 - 50 times higher than in plasma. Unlike plasma insulin levels, which fluctuate in response to meals, measurement of the 24-hour urinary excretion of C-Peptide provides a useful monitor of average beta cell insulin secretion.

The level of C-Peptide has been used to assess the following clinical applications:
  1. to assess the residual beta cell function in patients treated with insulin and to distinguish between types 1 and 2 diabetes. Of particular interest is its use to indicate the need for progression to insulin therapy in type 2 diabetes.
  2. The diagnosis of factitious hypoglycemia. The surreptitious administration of insulin causes high insulin levels in the absence of elevated C-Peptide concentrations.
  3. Insulinoma diagnosis, especially in patients treated with insulin. C-Peptide measurement is used in insulin suppression tests in euglycemic patients with suspected insulinoma. Elevated C-Peptide levels in this test are indicative of insulinoma.
  4. As a marker for residual pancreatic tissue after pancreatectomy. In the case of insulinoma, C-Peptide measurement may be used to detect metastasis and the response to therapy. It may also be used to monitor the progress of pancreas or islet cell transplantation

Product Features

  • Strip plates and additional reagents allow for use in multiple experiments
  • Quantitative protein detection
  • Establishes normal range
  • The best products for confirmation of antibody array data

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Standard Peptide
  • Assay Diluent(s)
  • Biotinylated Peptide
  • HRP-Streptavidin
  • TMB One-Step Substrate
  • Stop Solution
  • Assay Diagram
  • Positive Control Sample
  • Capture Antibody
  • User Manual
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 ml volumes
  • Adjustable 1-25 ml pipettes for reagent preparation
  • 100 ml and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Orbital shaker
  • Aluminum foil
  • Saran Wrap
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • SigmaPlot software (or other software that can perform four-parameter logistic regression models)
Protocol Outline
  • Prepare all reagents, samples and standards as instructed.
  • Add 100 µl detection antibody to each well.
  • Incubate 1.5 h at RT or O/N at 4°C.
  • Add 100 µl standard or sample to each well.
  • Incubate 2.5 h at RT.
  • Add 100 µl prepared streptavidin solution.
  • Incubate 45 min at RT.
  • Add 100 µl TMB One-Step Substrate Reagent to each well.
  • Incubate 30 min at RT.
  • Add 50 µl Stop Solution to each well.
  • Read plate at 450 nm immediately.


Standard, Biotinylated C-Peptide peptide, and Positive Control should be stored at -20°C after arrival. Avoid multiple freeze-thaws. The remaining kit components may be stored at 4°C. Opened Microplate Wells and antibody (Item N) may be stored for up to 1 month at 2° to 8°C. Return unused wells to the pouch containing desiccant pack and reseal along entire edge.

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This product is furnished for LABORATORY RESEARCH USE ONLY.
Not for diagnostic or therapeutic use.

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