Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human FLRG 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
Cell Culture Supernates (100 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL), Urine (50 uL), Human Milk (50 uL)
Sensitivity
8.55 pg/mL
Assay Range
62.5 - 4,000 pg/mL
Specificity
Natural and recombinant human FLRG
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 FLRG Immunoassay is a 4.5 hour solid-phase ELISA designed to measure human FLRG in cell culture supernates, serum, plasma, urine, and human milk. It contains NS0-expressed recombinant human FLRG and has been shown to accurately quantitate the recombinant factor. Results obtained using natural human FLRG 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 FLRG.
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 | 20 | 20 | 20 |
| Mean (pg/mL) | 468 | 1113 | 2252 | 494 | 1174 | 2331 |
| Standard Deviation | 10.4 | 23 | 55.2 | 32.9 | 69 | 143 |
| CV% | 2.2 | 2.1 | 2.5 | 6.7 | 5.9 | 6.1 |
Serum, EDTA Plasma, Heparin Plasma, Human Milk
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 448 | 1094 | 2306 | 486 | 1148 | 2282 |
| Standard Deviation | 15.4 | 18.5 | 49.5 | 29.5 | 53 | 122 |
| CV% | 3.4 | 1.7 | 2.1 | 6.1 | 4.6 | 5.3 |
Recovery
The recovery of FLRG spiked to levels throughout the range of the assay was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 107 | 98-111 |
| EDTA Plasma (n=4) | 96 | 85-105 |
| Heparin Plasma (n=4) | 102 | 89-114 |
| Serum (n=4) | 100 | 91-112 |
Linearity
To assess the linearity of the assay, samples containing high concentrations of FLRG were serially diluted with the appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay.
Scientific Data
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: Follistatin-related Gene Protein/FLRG
Follistatin-related gene protein (FLRG) has been shown to bind and inhibit the TGF-beta family ligands including activin, BMP-2, -6, -7, and GDF-8. FLRG is expressed most highly in heart, lung, kidney, and placenta, and is upregulated by TGF-beta and Activin via Smad proteins.
Entrez Gene IDs:
10272 (Human); 83554 (Mouse)
Alternate Names:
FLRG; FLRGFSRP; follistatin-like 3 (secreted glycoprotein); Follistatin-like 3; Follistatin-like protein 3; Follistatin-related gene protein; follistatin-related protein 3; FSTL3
⚠ 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 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.
- 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.
100 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Citations for Human FLRG Quantikine ELISA Kit
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.
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Citations: Showing 1 - 7
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The circulating proteomic signature of alcohol-associated liver disease
Authors: J Luther, AG Vannier, EA Schaefer, RP Goodman
JCI Insight, 2022-07-22;7(14):.
Species: Human
Sample Types: Serum
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The Activin/FLRG pathway associates with poor COVID-19 outcomes in hospitalized patients
Authors: M McAleavy, Q Zhang, PJ Ehmann, J Xu, MF Wipperman, D Ajithdoss, L Pan, M Wakai, R Simonson, A Gadi, A Oyejide, SC Hamon, A Boyapati, LG Morton, T Shavlakadz, CA Kyratsous, DJ Glass
Molecular and Cellular Biology, 2021-11-01;0(0):MCB0046721.
Species: Human
Sample Types: Cell Culture Supernates
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The RNA landscape of the human placenta in health and disease
Authors: S Gong, F Gaccioli, J Dopierala, U Sovio, E Cook, PJ Volders, L Martens, PDW Kirk, S Richardson, GCS Smith, DS Charnock-J
Nature Communications, 2021-05-11;12(1):2639.
Species: Human
Sample Types: Serum
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Activin subfamily peptides predict chronological age in humans
Authors: LV Barrios-Si, M Parnell, ZB Shinwari, GA Chaudhary, T Xenofontos, A van Bekhov, S McArthur, BT Elliott
Physiol Rep, 2018-09-01;6(17):e13823.
Species: Human
Sample Types: Plasma
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Serum biomarker for diagnostic evaluation of pulmonary arterial hypertension in systemic sclerosis
Authors: LM Rice, JC Mantero, EA Stratton, R Warburton, K Roberts, N Hill, RW Simms, R Domsic, HW Farber, R Layfatis
Arthritis Res. Ther., 2018-08-16;20(1):185.
Species: Human
Sample Types: Serum
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Vitamin D supplementation lowers thrombospondin-1 levels and blood pressure in healthy adults
Authors: AT Amaraseker, B Assadi-Kha, S Liu, M Black, G Dymmott, NM Rogers, AL Sverdlov, JD Horowitz, DTM Ngo
PLoS ONE, 2017-05-10;12(5):e0174435.
Species: Human
Sample Types: Plasma
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Serum activin A and follistatin levels in gestational diabetes and the association of the Activin A-Follistatin system with anthropometric parameters in offspring.
Authors: Naf S, Escote X, Ballesteros M, Yanez R, Simon-Muela I, Gil P, Albaiges G, Vendrell J, Megia A
PLoS ONE, 2014-04-24;9(4):e92175.
Species: Human
Sample Types: Serum
FAQs
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Human FLRG Quantikine ELISA Kit
By Anonymous
on 02/18/2021
Sample Tested: Serum
