Human Proinsulin Quantikine ELISA Kit

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DPINS0
Control Products Available
Human Proinsulin ELISA Standard Curve
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Human Proinsulin 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
Serum (100 uL), EDTA Plasma (100 uL), Heparin Plasma (100 uL)
Sensitivity
1.43 pM
Assay Range
3.3 - 210 pM (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Specificity
Natural and recombinant human Proinsulin.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
Interference observed with 1 or more available related molecules.

Product Summary

The Quantikine Human Proinsulin Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human Proinsulin in serum and plasma. It contains E. coli-expressed recombinant human Proinsulin and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human Proinsulin 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 human Proinsulin.

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

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 21.1 62.7 119 18 55.1 109
Standard Deviation 0.44 0.745 1.4 1.9 3.97 6.91
CV% 2.1 1.2 1.2 10.6 7.2 6.3

Recovery

The recovery of human Proinsulin spiked to levels throughout the range of the assay in various matrices was evaluated.

Sample Type Average % Recovery Range %
EDTA Plasma (n=4) 92 86-108
Heparin Plasma (n=4) 94 90-100
Serum (n=4) 95 80-105

Linearity

To assess the linearity of the assay, samples spiked with high concentrations of human Proinsulin were diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Human Proinsulin ELISA Linearity

Data Examples

Human Proinsulin ELISA Standard Curve

Product Datasheets

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: Proinsulin

Insulin is a peptide hormone that is critical for glucose homeostasis. The mature Insulin peptide is derived from Proinsulin, which includes the Insulin A and B chains connected by a peptide fragment (Insulin C-peptide). Proinsulin is processed within the endoplasmic reticulum of pancreatic beta cells into equimolar ratios of mature Insulin and Insulin C-peptide. Proinsulin binds the Insulin Receptor A isoform (IR-A) with high affinity resulting in cell proliferation and migration. It has low binding affinity for the Insulin Receptor B isoform (IR-B) and the Insulin-like Growth Factor I Receptor (IGF R1).

Entrez Gene IDs:
3630 (Human); 16333 (Mouse)
Alternate Names:
IDDM2; ILPR; insulin; IRDN; MODY10; Proinsulin
⚠ 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.

  3. 100 µL Assay Diluent
  4.   Add 100 µL of Assay Diluent to each well.

  5. 100 µL Standard, Control, or Sample
  6.   Add 100 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.
  7.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.

  8. 200 µL Conjugate
  9.   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.
  10.   Aspirate and wash 4 times.

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

  13. 50 µL Stop Solution
  14.   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.

FAQs

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