Human Proinsulin Quantikine ELISA Kit Summary
Serum, EDTA Plasma, Heparin Plasma
|Intra-Assay Precision||Inter-Assay Precision|
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|
Human Proinsulin ELISA Standard Curve
Preparation and Storage
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).
- Prepare all reagents, standard dilutions, and samples as directed in the product insert.
- Remove excess microplate strips from the plate frame, return them to the foil pouch containing the desiccant pack, and reseal.
- Add 100 µL of Assay Diluent to each well.
- 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.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- 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.
- Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- 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.
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