Overview

  • Product name

    Proinsulin ELISA kit
  • Detection method

    Colorimetric
  • Precision

    Intra-assay
    Sample n Mean SD CV%
    1 20 26.3pg/ml 5.5%
    2 20 9.6pg/ml 6.1%
    3 20 3.7pg/ml 17.4%
    Inter-assay
    Sample n Mean SD CV%
    1 26.1pg/ml 6%
    2 9pg/ml 6.3%
    3 3.3pg/ml 11.6%
  • Sample type

    Serum, EDTA Plasma
  • Assay type

    Sandwich (quantitative)
  • Sensitivity

    0.17 pmol/L
  • Range

    1.6 pmol/L - 50 pmol/L
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 94.6 72.3% - 111.3%
    EDTA Plasma 88.7 72.9% - 118.4%

  • Assay time

    3h 00m
  • Assay duration

    Multiple steps standard assay
  • Species reactivity

    Reacts with: Human
  • Product overview

    The Proinsulin ELISA Kit (ab213977) (in vitro Enzyme-Linked Immunosorbent Assay) is designed for the quantitative measurement of proinsulin in human serum and plasma.


    The ELISA kit uses a monoclonal antibody specific to the proinsulin C-peptide immobilized on a microtiter plate. Standards or samples containing proinsulin are added to the plate. After a short incubation excess sample or standard is washed out and a biotinylated monoclonal antibody to insulin is added. This antibody binds to the Proinsulin captured on the plate. After a short incubation excess antibody is washed out and Streptavidin conjugated to Horseradish peroxidase (SA-HRP) is added, which binds to the biotin on the bound monoclonal antibody. The plate is then incubated. Excess conjugate is washed out and TMB substrate solution is added and incubated. An HRP-catalyzed reaction generates a blue color in the solution. Stop solution is added to stop the substrate reaction. The resulting yellow color is read at 450nm. The amount of signal is directly proportional to the level of Proinsulin in the sample.

  • Notes

    Type 2 diabetes mellitus is the most common serious metabolic condition in the world, and results from a subnormal response of tissues to insulin (insulin resistance) and a failure of the insulin-secreting beta cells to compensate1Insulin is required for glucose homeostasis and an important precursor protein to insulin is proinsulin. Proinsulin is composed of the A and B subunits of insulin which is connected by the C-peptide region. This 10.5 kDa protein, comprised of 110 amino acids, is synthesized as a single chain that contains a 24 amino acid signal sequence and an 86 amino acid proinsulin propeptide. Proinsulin is processed in the endoplasmic reticulum of pancreatic β-cells and is converted within immature secretory granules into insulin and C-peptide. While mature insulin is required for glucose homeostasis and C-peptide is associated with renal function, proinsulin has low metabolic activity. However, it is biologically active and is a regulator of development in pre-pancreatic embryonic stages.

    The release of proinsulin is significantly increased in patients with type 2 diabetes as well as in individuals with impaired glucose tolerance (IGT). This increase in diabetic patients is commonly referred to as hyperproinsulinaemia and could result from a defect in the processing of proinsulin to mature insulin. This defect is an early sign of an abnormality of pancreatic β cell dysfunction and results in elevated proinsulin:insulin ratios. The levels of proinsulin have also been found to increase with age.

  • Tested applications

    Suitable for: Sandwich ELISAmore details
  • Platform

    Pre-coated microplate (12 x 8 well strips)

Properties

  • Storage instructions

    Store at +4°C. Please refer to protocols.
  • Components 1 x 96 tests
    Antibody Diluent 1 x 11ml
    Anti-Proinsulin coated microplate (12x 8 well strips) 1 unit
    Plate Sealer 3 units
    Proinsulin Antibody (Lyophilized) 1 vial
    Proinsulin Conjugate 1 x 20ml
    Proinsulin Standard (10.5 ng/ml) 1 vial
    Sample Diluent 1 x 20ml
    Stop Solution (1N Soln.) 1 x 10ml
    TMB Substrate 1 x 10ml
    Wash Buffer Concentrate 1 x 100ml
  • Research areas

  • Relevance

    A precursor of insulin it is converted to insulin by removal of the connecting C peptide, leaving the two (A and B) chains. From every molecule of proinsulin, one molecule of insulin plus one molecule of C Peptide are produced.
  • Cellular localization

    Secreted
  • Alternative names

    • INS
    • insulin
    • IRDN
    • Proinsulin precursor
  • Database links

Applications

Our Abpromise guarantee covers the use of ab213977 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
Sandwich ELISA Use at an assay dependent concentration.

Images

Protocols

References

ab213977 has not yet been referenced specifically in any publications.

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