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Human/Canine/Porcine Insulin ELISA Kit - Quantikine Best Seller

R&D Systems, part of Bio-Techne | Catalog # DINS00

R&D Systems, part of Bio-Techne
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DINS00

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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)

Sensitivity

2.15 pmol/L

Assay Range

15.6-500 pmol/L (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)

Product Summary for Human/Canine/Porcine Insulin Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Reactivity

Human, Porcine, Canine

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.


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, EDTA Plasma, Heparin Plasma, Serum

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 4.60 10.2 21.5
CV% 3.9 3.7 4.0 7.4 6.9 7.5

Recovery for Human/Canine/Porcine Insulin Quantikine ELISA Kit

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 Images for Human/Canine/Porcine Insulin Quantikine ELISA Kit

Human/Porcine/Canine Insulin ELISA Standard Curve

Human/Porcine/Canine Insulin ELISA Standard Curve

Preparation and Storage

Shipping

The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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.

Alternate Names

IDDM2, ILPR, INS, IRDN, MODY10

Entrez Gene IDs

3630 (Human); 16333 (Mouse); 397415 (Porcine); 280829 (Bovine); 483665 (Canine)

Gene Symbol

INS

Additional Insulin Products

Product Documents for Human/Canine/Porcine Insulin Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human/Canine/Porcine Insulin Quantikine ELISA Kit

For research use only

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⚠ 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.