Human AgRP Quantikine ELISA Kit

  (2 citations)     
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Assay Procedure
Citations (2)
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  • 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), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (25 uL)
  • Sensitivity
    2.68 pg/mL
  • Assay Range
    7.8 - 500 pg/mL (Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Urine)
  • Specificity
    Natural and recombinant human AgRP
  • Cross-reactivity
    Cross-reactivity observed with 1 or more available related molecules.< 50% cross-species reactivity observed with species tested.
  • Interference
    No significant interference observed with available related molecules.
Control Available
QC24 , Quantikine Immunoassay Control Group 9 - Please Inquire
Product Summary
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.

Precision
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
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 54.3 143 257 43.7 126 238
Standard Deviation 2.64 3.88 5.69 3.91 8.24 16.3
CV% 4.9 2.7 2.2 8.9 6.5 6.8

EDTA Plasma, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 40 40 40
Mean 56.6 144 262 45.1 130 246
Standard Deviation 3.14 4.86 7.82 4.66 10 18.1
CV% 5.5 3.4 3 10.3 7.7 7.4

Recovery

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

Sample Type Average % Recovery Range %
Cell Culture Media (n=4) 106 95-115
EDTA Plasma (n=4) 96 89-106
Heparin Plasma (n=4) 92 84-103
Linearity
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.
 AgRP/ART [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Preparation and Storage
  • Storage
    Store the unopened product at 2 - 8 °C. Do not use past expiration date.
Background: AgRP/ART

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.

  • Long Name:
    Agouti-related Protein
  • Entrez Gene IDs:
    181 (Human); 11604 (Mouse)
  • Alternate Names:
    agouti related protein homolog (mouse); agouti-related protein; AgRP; Agrt; AGRTMGC118963; ART; ARTagouti (mouse) related protein; ASIP2
Related Research Areas
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. 50 µL Standard, Control, or Sample
  6.   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.
  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.
Citations:

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.

2 Citations: Showing 1 - 2
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Species
Sample Type
  1. Effects of estradiol on cerebrospinal fluid levels of agouti-related protein in ovariectomized rhesus monkeys.
    Authors: Xiao E, Kim AJ, Dutia R
    Endocrinology, 2010;151(3):1002-9.
    Species: Primate - Macaca mulatta (Rhesus Macaque)
    Sample Type: Tissue Homogenates
  2. Plasma agouti-related protein levels in women with anorexia nervosa.
    Authors: Moriya J, Takimoto Y, Yoshiuchi K
    Psychoneuroendocrinology, 2006;31(9):1057-61.
    Species: Human
    Sample Type: Plasma

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Quantikine Immunoassay Control Group 9

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Quantikine ELISA Wash Buffer 1

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