The Quantikine Human AgRP Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human AgRP in cell culture supernates, plasma, and urine. It contains Sf 21-expressed recombinant human AgRP and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human AgRP 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 AgRP.
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested on one plate to assess intra-assay precision
Inter-Assay Precision (Precision between assays) Three samples of known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Urine
EDTA Plasma, Heparin Plasma
The recovery of AgRP spiked to levels throughout the range of the assay in various matrices was evaluated.
Average % Recovery
Cell Culture Media (n=4)
EDTA Plasma (n=4)
Heparin Plasma (n=4)
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of AgRP were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Agouti-related Protein (AgRP), also known as the protein product of the Agouti-related Transcript (ART), is a neuropeptide and a functional antagonist of alpha-MSH on Melanocortin (MC)-3 and MC-4 receptors, both of which are implicated in obesity. AgRP shares sequence and structural similarity with the Agouti protein and has similar expression patterns in both mouse and human.
Entrez Gene IDs:
181 (Human); 11604 (Mouse)
agouti related protein homolog (mouse); agouti-related protein; AgRP; Agrt; AGRTMGC118963; ART; ARTagouti (mouse) related protein; ASIP2
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.
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.
100 µL Assay Diluent
Add 100 µL of Assay Diluent to each well.
50 µL Standard, Control, or Sample
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.
Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
200 µL Conjugate
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.
200 µL Substrate Solution
Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
50 µL Stop Solution
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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