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Human, Canine, & Porcine Insulin ELISA Kit - Quantikine

Catalog #: DINS00
17 Citations 3 Reviews Datasheet / COA / SDS
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Human/Porcine/Canine Insulin ELISA Standard Curve
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Summary Sample Values Precision Recovery Linearity Data Examples Product Datasheets Preparation and Storage Background Related Research Areas

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.

Sample Values

Serum/Plasma - Samples from apparently healthy, fasting volunteers were evaluated for the presence of human/canine/porcine Insulin in this assay. No medical histories were available for the donors used in this study.

Human SamplesMean (pmol/L)Range (pmol/L)Standard Deviation (pmol/L)
Serum (n=34)58.017.2-21841
EDTA plasma (n=34)60.315.8-20839
Heparin plasma (n=34)59.416.4-19045

Canine SamplesMean (pmol/L)Range (pmol/L)Standard Deviation (pmol/L)
Serum (n=10)58.816.7-19052
Heparin plasma (n=5)78.321.5-14656

Canine Samples Mean of Detectable (pmol/L)% DetectableRange  (pmol/L)
EDTA plasma (n=5)96.480ND-145
ND=Non-detectable

Porcine SamplesMean of Detectable (pmol/L)% DetectableRange (pmol/L)
Serum (n=10)22.690ND-34.3
EDTA plasma (n=5)27.180ND-40.8
Heparin plasma (n=5)30.480ND-40.9
ND=Non-detectable

Cell Culture Supernates - Human peripheral blood mononuclear cells (PBL) were cultured in DMEM supplemented with 5% fetal bovine serum, 50 μM beta -mercaptoethanol, 2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate. Cells were cultured unstimulated or stimulated with 10 μg/mL PHA for 1 and 5 days. Aliquots of the cell culture supernates were removed and assayed for human/canine/porcine Insulin. No detectable levels were observed.


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.

Precision

Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in twenty separate assays to assess inter-assay precision. Assays were performed by at least three technicians using two lots of components

Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean (pmol/L) 66.8 158 299 62.2 148 288
Standard Deviation 2.58 5.89 11.9 4.6 10.2 21.5
CV% 3.9 3.7 4 7.4 6.9 7.5

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

N/A Human/Porcine/Canine Insulin ELISA Standard Curve View Larger

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
    2.   Add 150 µL of Assay Diluent to each well.
      3. 50 µL Standard, Control, or Sample
    3.   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.
    4.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
      5. 200 µL Conjugate
    5.   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.
    6.   Aspirate and wash 4 times.
      7. 200 µL Substrate Solution
    7.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
      8. 50 µL Stop Solution
    8.   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.

17 Citations: Showing 1 - 10
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  1. Rapid benefits in older age from transition to whole food diet regardless of protein source or fat to carbohydrate ratio: Arandomised control trial
    Authors: Ribeiro, RV;Senior, AM;Simpson, SJ;Tan, J;Raubenheimer, D;Le Couteur, D;Macia, L;Holmes, A;Eberhard, J;O'Sullivan, J;Koay, YC;Kanjrawi, A;Yang, J;Kim, T;Gosby, A;
    Aging cell
    Species: Human
    Sample Types: Plasma
  2. Effects of high-intensity interval training on cardiometabolic biomarkers in patients with prostate cancer undergoing active surveillance: a randomized controlled trial
    Authors: Kang, DW;Field, CJ;Patel, D;Fairey, AS;Boulé, NG;Dieli-Conwright, CM;Courneya, KS;
    Prostate cancer and prostatic diseases
    Species: Human
    Sample Types: Serum
  3. Linking adipose tissue eosinophils, interleukin-4, and leptin in human obesity and insulin resistance
    Authors: Hernandez, JD;Li, T;Ghannam, H;Rau, CM;Masuda, MY;Madura, JA;Jacobsen, EA;Filippis, E;
    JCI insight
    Species: Human
    Sample Types: Serum, Plasma
  4. Feasibility and metabolic outcomes of a well-formulated ketogenic diet as an adjuvant therapeutic intervention for women with stage IV metastatic breast cancer: The Keto-CARE trial
    Authors: Buga, A;Harper, DG;Sapper, TN;Hyde, PN;Fell, B;Dickerson, R;Stoner, JT;Kackley, ML;Crabtree, CD;Decker, DD;Robinson, BT;Krystal, G;Binzel, K;Lustberg, MB;Volek, JS;
    PloS one
    Species: Human
    Sample Types:
  5. 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
  6. 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
  7. 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
  8. 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
  9. 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
  10. 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
  11. 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
  12. 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
  13. 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
  14. 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
  15. 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
  16. 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
  17. 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

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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 DINS00
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.

Human/Canine/Porcine Insulin Quantikine ELISA Kit DINS00