Human Insulin ELISA

RayBio® Human Insulin ELISA Kit for cell culture supernatants, plasma, and serum samples.

ELH-Insulin-1
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Antigen Information

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

Assay Format

Detection Range:
4 uIU/ml - 300 uIU/ml
Recommended Dilution (serum & plasma):
2 - 10 fold
Sensitivity:
4 uIU/ml

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Specificity:
This ELISA kit shows no cross-reactivity with the following cytokines tested: human BDNF, BLC, ENA-78, IL-1 alpha, IL-1 beta, IL-2, IL-3, IL-5, IL-6, IL-7, IL-8, IL-9, IL-11, IL-12 p70, IL-12 p40, IL-13, IL-15, I-309, IP-10, G-CSF, GM-CSF, IFN-gamma, Leptin (OB), MCP-1, MCP-2, MCP-3, MDC, MIP-1 alpha, MIP-1 beta, MIP-1 delta, PARC, PDGF, RANTES, SCF, TARC, TGF-beta, TIMP-1, TNF-alpha, TNF-beta, TPO, VEGF.
Number of Targets Detected:
1
Species Detected:
  • Human
Compatible Sample Types:
  • Cell Culture Supernatants
  • Plasma
  • Serum
Design Principle:
  • Sandwich-based
Method of Detection:
Colorimetric
Quantitative/Semi-Quantitative:
Quantitative
Solid Support:
96-well Microplate

Product Specifications

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

Product Features

Additional Notes
  • 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

Standard Curves

Spiking & Recovery Results

Linearity Results

Application Notes

Kit Components
  • Pre-Coated 96-well Strip Microplate
  • Wash Buffer
  • Stop Solution
  • Assay Diluent(s)
  • Lyophilized Standard
  • Biotinylated Detection Antibody
  • Streptavidin-Conjugated HRP
  • TMB One-Step Substrate
Other Materials Required
  • Distilled or deionized water
  • Precision pipettes to deliver 2 µl to 1 µl volumes
  • Adjustable 1-25 µl pipettes for reagent preparation
  • 100 µl and 1 liter graduated cylinders
  • Tubes to prepare standard and sample dilutions
  • Absorbent paper
  • Microplate reader capable of measuring absorbance at 450nm
  • Log-log graph paper or computer and software for ELISA data analysis
Protocol Outline
  1. Prepare all reagents, samples and standards as instructed in the manual.
  2. Add 100 µl of standard or sample to each well.
  3. Incubate 2.5 h at RT or O/N at 4°C.
  4. Add 100 µl of prepared biotin antibody to each well.
  5. Incubate 1 h at RT.
  6. Add 100 µl of prepared Streptavidin solution to each well.
  7. Incubate 45 min at RT.
  8. Add 100 µl of TMB One-Step Substrate Reagent to each well.
  9. Incubate 30 min at RT.
  10. Add 50 µl of Stop Solution to each well.
  11. Read at 450 nm immediately.

Storage/Stability

The entire kit may be stored at -20°C for up to 1 year from the date of shipment. Avoid repeated freeze-thaw cycles. The kit may be stored at 4°C for up to 6 months. For extended storage, it is recommended to store at -80°C.
  1. Townsend J., Joffman J., Gonzalez A., et al. Effects of ?-Hydroxy-?-methylbutyrate Free Acid Ingestion and Resistance Exercise on the Acute Endocrine Response. Int J Endocrinol. 2015;2015:856708. doi: 10.1155/2015/856708.
    Species: Human
    Sample type: Serum
  2. Fadoua G., Cabrera A., Ferid M., et al. Leptin, Insulin and Lipid Profiles in Obese Subjects with and without Metabolic Syndrome in the Region of Cap-Bon: Tunisia. Endocrinol Metab Synd 2015, 4:3 http://dx.doi.org/10.4172/2161-1017.1000193
    Species: Human
    Sample type: Serum
  3. Agrawal A., Urimi D., Harde H., et al. Folate appended chitosan nanoparticles augment the stability, bioavailability and efficacy of insulin in diabetic rats following oral administration. RSC Adv., 2015,5, 105179-105193 DOI: 10.1039/C5RA19115G
    Species: Human
    Sample type: Plasma
  4. Jorgensen A., Nellemann C., Wohlfahr-Veje C. Interaction between paraoxonase 1 polymorphism and prenatal pesticide exposure on metabolic markers in children using a multiplex approach. Reprod Toxicol. 2014 Nov 22;51C:22-30. doi: 10.1016/j.reprotox.2014.11.005.
    Species: Human
    Sample type: Serum
  5. Leisegang K., Bouic P., Menkveld R., Henkel R. Reproductive Biology and Endocrinology 2014, 12:34 http://www.rbej.com/content/12/1/34
    Species: Human
    Sample type: Conditioned Media
  6. Rathore S., Bidwai A. Association of Insulin Resistance and Selected Adipokines in Cardiovascular Disease with and without Metabolic Syndrome. JMSCR Volume 2, Issue 5, Pages 1176-1186, May 2014
    Species: Human
    Sample type: Serum
  7. Monteiro PA, Mota J, Silveira LS, et al. Morphological and metabolic determinants of nonalcoholic fatty liver disease in obese youth: a pilot study. BMC Res Notes. 2013;6:89.
    Species: Human
    Sample type: Serum
  8. Silveira, Loreana Sanches, et al. "Intra-abdominal fat is related to metabolic syndrome and non-alcoholic fat liver disease in obese youth." BMC pediatrics 13.1 (2013): 115.
    Species: Human
    Sample type: Plasma
  9. Ching C., Sun T., Huang W., et al. A circuit design of a low-cost, portable and programmable electroporation device for biomedical applications. Sensors and Actuators B 166- 167 (2012) 292- 300
    Species: Human
    Sample type: Purified Protein
  10. Kumar R., Prakash S., Chhabra S., et al. Association of pro-inflammatory cytokines, adipokines & oxidative stress with insulin resistance & non-alcoholic fatty liver disease. Indian J Med Res. 2012 Aug;136(2):229-36.
    Species: Human
    Sample type: Serum
This kit was easy to use with very clear instructions.
Assay time is ~5 hours, but with washes and sample dispensing, it will take about 6.5 hours if done manually.
Snap apart wells allow you to customize how many assays you can run at once, but the "rack" they snap into weakens over time, so about the 2nd or 3rd use you may find the wells fall out very easily.
Only issue I had was the top 2 concentrations of the standard curve maxed out the signal, so be sure to dilute your samples to be sure they are not in the upper range of the assay.
This kit has worked well twice in my hands. The instructions are easy to interpret, and the detection is as expected. I plan to repurchase in the future
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Usage

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