Overview

  • Product name
    Human Insulin ELISA Kit, Fluorescent
  • Detection method
    Fluorescent
  • Precision
    Intra-assay
    Sample n Mean SD CV%
    Serum 5 8.6%
    Inter-assay
    Sample n Mean SD CV%
    Serum 3 4.9%
  • Sample type
    Serum, Heparin Plasma
  • Assay type
    Sandwich (quantitative)
  • Sensitivity
    1 pmol/L
  • Range
    1 pmol/L - 267.2 pmol/L
  • Recovery

    Sample specific recovery
    Sample type Average % Range
    Serum 106 103% - 111%
    Heparin Plasma 112 108% - 120%

  • Assay time
    1h 30m
  • Assay duration
    One step assay
  • Species reactivity
    Reacts with: Human
    Does not react with: Sheep, Goat, Guinea pig, Dog
  • Product overview

    Insulin in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Insulin protein in human serum and plasma (heparin).


    This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
    If using a Molecular Devices’ plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.


    The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

  • Notes

    Insulin is a highly conserved, secreted hormone essential for glucose metabolism. Produced by pancreatic beta cells, proinsulin is proteolyzed into an A and a B chain, which form a 6 kDa mature protein. Basal levels of insulin are continuously delivered into the bloodstream, and additional levels are secreted proportional to food ingestion. Insulin secretion is highly regulated, and dysregulation of insulin production or sensitivity results in Type 1 diabetes mellitus or Type 2 diabetes mellitus, respectively.

  • 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
    100X Stoplight Red Substrate 1 x 120µl
    10X Human Insulin Capture Antibody 1 x 600µl
    10X Human Insulin Detector Antibody 1 x 600µl
    10X Wash Buffer PT (ab206977) 1 x 20ml
    500X Hydrogen Peroxide (H2O2, 3%) 1 x 50µl
    Antibody Diluent 5BI 1 x 6ml
    Human Insulin Lyophilized Recombinant Protein 2 vials
    Plate Seals 1 unit
    Sample Diluent NS 1 x 50ml
    SimpleStep Pre-Coated Black 96-Well Microplate 1 unit
    Stoplight Red Substrate Buffer 1 x 12ml
  • Research areas
  • Function
    Insulin decreases blood glucose concentration. It increases cell permeability to monosaccharides, amino acids and fatty acids. It accelerates glycolysis, the pentose phosphate cycle, and glycogen synthesis in liver.
  • Involvement in disease
    Defects in INS are the cause of familial hyperproinsulinemia (FHPRI) [MIM:176730].
    Defects in INS are a cause of diabetes mellitus insulin-dependent type 2 (IDDM2) [MIM:125852]. IDDM2 is a multifactorial disorder of glucose homeostasis that is characterized by susceptibility to ketoacidosis in the absence of insulin therapy. Clinical fetaures are polydipsia, polyphagia and polyuria which result from hyperglycemia-induced osmotic diuresis and secondary thirst. These derangements result in long-term complications that affect the eyes, kidneys, nerves, and blood vessels.
    Defects in INS are a cause of diabetes mellitus permanent neonatal (PNDM) [MIM:606176]. PNDM is a rare form of diabetes distinct from childhood-onset autoimmune diabetes mellitus type 1. It is characterized by insulin-requiring hyperglycemia that is diagnosed within the first months of life. Permanent neonatal diabetes requires lifelong therapy.
    Defects in INS are a cause of maturity-onset diabetes of the young type 10 (MODY10) [MIM:613370]. MODY10 is a form of diabetes that is characterized by an autosomal dominant mode of inheritance, onset in childhood or early adulthood (usually before 25 years of age), a primary defect in insulin secretion and frequent insulin-independence at the beginning of the disease.
  • Sequence similarities
    Belongs to the insulin family.
  • Cellular localization
    Secreted.
  • Information by UniProt
  • Alternative names
    • IDDM
    • IDDM1
    • IDDM2
    • ILPR
    • ins
    • INS_HUMAN
    • Insulin A chain
    • Insulin B chain
    • IRDN
    • MODY10
    • Preproinsulin
    • Proinsulin
    • Proinsulin precursor
    see all
  • Database links

Applications

Our Abpromise guarantee covers the use of ab229416 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

  • ELISA Protocol Summary
  • Background-subtracted data values (mean +/- SD) are graphed.

  • Data shown for serum is from an individual donor with high endogenous insulin levels. Data shown for plasma is from a pooled (n=50) sample. Background-subtracted data values (mean +/- SD, n = 3) are graphed.

  • Ten individual healthy donors were evaluated for the presence of Insulin in serum using this assay. The range was from 11.8 – 345.1 pmol/L, with an average of 99.4 pmol/L. Health history and dietary status of donors were unknown.

Protocols

References

ab229416 has not yet been referenced specifically in any publications.

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