Human/Canine/Porcine Insulin Quantikine ELISA Kit

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Human/Porcine/Canine Insulin ELISA Standard Curve
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Human/Canine/Porcine Insulin Quantikine ELISA Kit Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL)
Sensitivity
2.15 pmol/L
Assay Range
15.6 - 500 pmol/L (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Recombinant human Insulin and natural human, canine, and porcine Insulin.  This assay cross-reacts 0.25% with Human Proinsulin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.Cross-species reactivity observed with 1 or more species tested.
Interference
No significant interference observed with available related molecules.

Product Summary

The Quantikine Human/Canine/Porcine Insulin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure Insulin in cell culture supernates, serum, and plasma. It contains S. cerevisiae-expressed recombinant human Insulin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural Insulin 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 Insulin.

Recovery

The recovery of Insulin spiked to levels throughout the range of the assay in various matrices was evaluated

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 95 85-108
EDTA Plasma (n=4) 95 86-102
Heparin Plasma (n=4) 92 85-105
Serum (n=4) 96 87-103

Linearity

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

Scientific Data

Human/Porcine/Canine Insulin ELISA Standard Curve

Product Datasheets

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Preparation and Storage

Shipping
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.
Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: Insulin

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.

Entrez Gene IDs:
3630 (Human); 16333 (Mouse); 397415 (Porcine); 280829 (Bovine); 483665 (Canine)
Alternate Names:
IDDM2; ILPR; INS; Insulin; IRDN; MODY10; proinsulin
&#9888; 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.
    1.   Aspirate each well and wash, repeating once more for a total of 2 washes.

    2. 150 µL Assay Diluent
    3.   Add 150 µL of Assay Diluent to each well.

    4. 50 µL Standard, Control, or Sample
    5.   Add 50 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours on a horizontal orbital microplate shaker.
    6.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

    7. 200 µL Conjugate
    8.   Add 200 µL of Conjugate to each well. Cover with a new plate sealer, and incubate at room temperature for 2 hours on the shaker.
    9.   Aspirate and wash 4 times.

    10. 200 µL Substrate Solution
    11.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

    12. 50 µL Stop Solution
    13.   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.

Citations for Human/Canine/Porcine Insulin Quantikine ELISA Kit

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

13 Citations: Showing 1 - 10
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  1. Is There a Correlation between Apelin and Insulin Concentrations in Early Second Trimester Amniotic Fluid with Fetal Growth Disorders?
    Authors: Vrachnis, D;Antonakopoulos, N;Fotiou, A;Pergialiotis, V;Loukas, N;Valsamakis, G;Iavazzo, C;Stavros, S;Maroudias, G;Panagopoulos, P;Vlahos, N;Peppa, M;Stefos, T;Mastorakos, G;
    Journal of clinical medicine
    Species: Human
    Sample Types: Amniotic Fluid
  2. Evaluation of safety and anti-obesity effects of DWP16001 in naturally obese dogs
    Authors: B Rhee, RM Mahbubur, C Jin, JS Choi, HW Lim, W Huh, JS Park, J Han, S Kim, Y Lee, J Park
    Bmc Veterinary Research, 2022-06-22;18(1):237.
    Species: Canine
    Sample Types: Serum
  3. Increased Fibroblast Growth Factor 21 (FGF21) Concentration in Early Second Trimester Amniotic Fluid and Its Association with Fetal Growth
    Authors: N Vrachnis, S Argyridis, D Vrachnis, N Antonakopo, G Valsamakis, C Iavazzo, D Zygouris, N Salakos, A Rodolakis, N Vlahos, G Mastorakos, P Drakakis, Z Iliodromit
    Metabolites, 2021-08-28;11(9):.
    Species: Human
    Sample Types: Amniotic Fluid
  4. Lifestyle Intervention for Overweight/Obese Pregnant Women with Polycystic Ovarian Syndrome: Lessons and Challenges
    Authors: C Liu, L Zhang, W Zheng, X Liang, L Zhang, Z Tian, G Li
    Obesity facts, 2021-07-26;14(4):405-414.
    Species: Human
    Sample Types: Plasma
  5. A Comparison of the Gluco-Regulatory Responses to High-Intensity Interval Exercise and Resistance Exercise
    Authors: BA Gordon, CJ Taylor, JE Church, SD Cousins
    International Journal of Environmental Research and Public Health, 2021-01-02;18(1):.
    Species: Human
    Sample Types: Plasma
  6. Effects of Pre-Sleep Whey vs. Plant-Based Protein Consumption on Muscle Recovery Following Damaging Morning Exercise
    Authors: PG Saracino, HE Saylor, BR Hanna, RC Hickner, JS Kim, MJ Ormsbee
    Nutrients, 2020-07-10;12(7):.
    Species: Human
    Sample Types: Plasma
  7. Study protocol to investigate biomolecular muscle profile as predictors of long-term urinary incontinence in women with gestational diabetes mellitus
    Authors: MVC Rudge, FP Souza, JF Abbade, RLS Hallur, JPC Marcondes, F Piculo, G Marini, G Vesentini, L Thabane, SS Witkin, IMP Calderon, AMP Barbosa, MV Rudge, AMP Barbosa, IMP Calderon, FP Souza, JF Abbade, LSR Hallur, F Piculo, G Marini, G Vesentini, L Thabane, MS Palma, CFO Graeff, RK Arni, RD Herculano, DF Salvadori, S Mateus, M Dal Pai Si, CG Magalhães, RA Costa, SAM Lima, SL Felisbino, W Barbosa, A Atallah, MJB Girão, Z Di Bella, SM Uchoa, S Payão, A Hijas, B Berghman, R De Bie, L Sobrevia, B Junginger, FCB Alves, PS Rossignoli, CB Prudencio, MIG Orlandi, MI Gonçalves, SK Nunes, BB Catinelli, S Quiroz, BV Sarmento, FA Pinheiro, CI Sartorão, RR Lucas, DRA Reyes, SBCV Quiroz, EMA Enriquez, RG Oliveira, JF Floriano, JPC Marcondes, S Barneze, TD Dangió, T Pascon, P Rossignoli, JV Freitas, L Takano, F Reis, TD Caldeirão, JN Fernandes, AM Carr, MVC Gaitero, JE Corrente, HRC Nunes, AF Candido, SMB Costa, TD Dangió, T Pascon, JVF Melo, L Takano, FVDS Reis, TD Caldeirão, AM Carr, GA Garcia, GB Rabadan, HCM Bassin, KS Suyama, LN Damasceno, MLS Takemoto, MD Menezes, DG Bussaneli, VKC Nogueira, PR Lima, IO Lourenço, J Marostica, RA Megid, IP Caruso, LT Rasmussen, GM Prata, F Piculo, G Vesentini, MA Arantes, GAR Ferraz, LP Camargo, MR Kron, JE Corrente, HRC Nunes
    BMC Pregnancy Childbirth, 2020-02-19;20(1):117.
    Species: Human
    Sample Types: Serum
  8. The Fragility of Cryopreserved Insulin-producing Cells Differentiated from Adipose-tissue-derived Stem Cells
    Authors: K Tokuda, T Ikemoto, Y Saito, K Miyazaki, S Yamashita, S Yamada, S Imura, Y Morine, M Shimada
    Cell Transplant, 2020-01-01;29(0):9636897209547.
    Species: Human
    Sample Types: Cell Culture Supernates
  9. A change in the zinc ion concentration reflects the maturation of insulin-producing cells generated from adipose-derived mesenchymal stem cells
    Authors: S Ohta, T Ikemoto, Y Wada, Y Saito, S Yamada, S Imura, Y Morine, M Shimada
    Sci Rep, 2019-12-10;9(1):18731.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. Phospholipid complex based nanoemulsion system for oral insulin delivery: preparation, in vitro, and in vivo evaluations
    Authors: XB Hu, TT Tang, YJ Li, JY Wu, JM Wang, XY Liu, DX Xiang
    Int J Nanomedicine, 2019-05-01;14(0):3055-3067.
    Species: Rat
    Sample Types: Plasma
  11. Porcine epidemic diarrhea virus reduces feed efficiency in nursery pigs
    Authors: SM Curry, ER Burrough, KJ Schwartz, KJ Yoon, SM Lonergan, NK Gabler
    J. Anim. Sci., 2018-02-15;0(0):.
    Species: Porcine
    Sample Types: Serum
  12. The Effects of 3 Weeks of Uphill and Downhill Walking on Blood Lipids and Glucose Metabolism in Pre-Diabetic Men: A Pilot Study
    Authors: M Philippe, H Gatterer, EM Eder, A Dzien, M Somavilla, A Melmer, C Ebenbichle, T Müller, M Burtscher
    J Sports Sci Med, 2017-03-01;16(1):35-43.
    Species: Human
    Sample Types: Plasma
  13. Enhanced leptin sensitivity, reduced adiposity, and improved glucose homeostasis in mice lacking exchange protein directly activated by cyclic AMP isoform 1.
    Authors: Yan J, Mei F, Cheng H, Lao D, Hu Y, Wei J, Patrikeev I, Hao D, Stutz S, Dineley K, Motamedi M, Hommel J, Cunningham K, Chen J, Cheng X
    Mol Cell Biol, 2012-12-21;33(5):918-26.
    Species: Mouse
    Sample Types: Plasma

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Reviews for Human/Canine/Porcine Insulin Quantikine ELISA Kit

Average Rating: 4.7 (Based on 3 Reviews)

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Human/Canine/Porcine Insulin Quantikine ELISA Kit
By Anonymous on 11/22/2021
Sample Tested: human floxp embryonal stem cell culture medium

Human/Canine/Porcine Insulin Quantikine ELISA Kit
By Anonymous on 10/25/2018
Sample Tested: Cell Culture Media

Human/Canine/Porcine Insulin Quantikine ELISA Kit
By Bradley Elliott on 04/12/2018
Sample Tested: Heparin Plasma

Ran here according to manufacturers instructions vs a Duoset Kit (#DY8056-05) before purchasing multiple kits for large study. Human heparinised plasma from an IVGTT, ran in both plates at the same time, by the same researcher on the same aliquots.

Broadly - Duoset signal was stronger throughout, and resultant IVGTT curve was smoother. Quantikine kit may be underexposed with 30 minutes substrate development in the Quantikine instructions? We've moved forward with the Duosets.