Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Human MFG-E8 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 (50 uL), Serum (20 uL), EDTA Plasma (20 uL), Heparin Plasma (20 uL), Saliva (50 uL), Urine (50 uL), Human Milk (10 uL)
Sensitivity
13.4 pg/mL
Assay Range
62.5 - 4,000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
Specificity
Natural and recombinant human MFG-E8.
Cross-reactivity
< 0.5% cross-reactivity observed with available related molecules.< 50% cross-species reactivity observed with species tested.
Interference
No significant interference observed with available related molecules.
Product Summary
The Quantikine Human MFG-E8 Immunoassay is a 4.5 hour solid phase ELISA designed to measure MFG-E8 in cell culture supernates, serum, plasma, saliva, urine, and human milk. It contains NS0-expressed recombinant human MFG-E8 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate recombinant human MFG-E8. Results obtained using natural human MFG-E8 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 natural human MFG-E8.
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 a 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 kit components
Cell Culture Supernates, Urine, 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) | 680 | 1181 | 2103 | 692 | 1262 | 2206 |
| Standard Deviation | 30.5 | 38.9 | 84.7 | 57.6 | 88 | 125 |
| CV% | 4.5 | 3.3 | 4 | 8.3 | 7 | 5.7 |
Serum, EDTA Plasma, Heparin Plasma, Saliva
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 645 | 1143 | 2168 | 749 | 1326 | 2339 |
| Standard Deviation | 39.5 | 46.6 | 81.1 | 57.4 | 78 | 117 |
| CV% | 6.1 | 4.1 | 3.7 | 7.7 | 5.9 | 5 |
Recovery
The recovery of human MFG-E8 spiked to levels throughout the range of the assay in various matrices was evaluated. Samples were diluted prior to assay.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=4) | 105 | 100-110 |
| EDTA Plasma (n=4) | 96 | 89-102 |
| Heparin Plasma (n=4) | 96 | 89-102 |
| Serum (n=4) | 101 | 92-110 |
Linearity
To assess the linearity of the assay, samples containing high
concentrations of MFG-E8 were serially diluted with the appropriate Calibrator Diluent to
produce samples with values within the dynamic range of the assay.
Samples were diluted prior to assay.
Scientific Data
Product Datasheets
Preparation and Storage
Shipping
The product is shipped with polar packs. 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: MFG-E8
MFG-E8, originally identified as a product of lactating mammary glands, is also secreted by activated macrophages and immature dendritic cells. MFG-E8 functions as an opsonin by binding phosphatidylserine on apoptotic cells and enhancing their phagocytosis.
Long Name:
Milk Fat Globule EGF Factor 8
Entrez Gene IDs:
4240 (Human); 17304 (Mouse)
Alternate Names:
BA46; Breast epithelial antigen BA46; EDIL1; hP47; HsT19888; lactadherin; Lactahedrin; Medin; MFG1; MFGE8; MFG-E8; MFGM; milk fat globule-EGF factor 8 protein; Milk fat globule-EGF factor 8; O-acetyl disialoganglioside synthase; OAcGD3S; SED1; SPAG10; sperm associated antigen 10; sperm surface protein hP47
⚠ 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 MFG-E8 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.
12
Citations: Showing 1 - 10
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Medin co-aggregates with vascular amyloid-beta in Alzheimer's disease
Authors: J Wagner, K Degenhardt, M Veit, N Louros, K Konstantou, A Skodras, K Wild, P Liu, U Obermüller, V Bansal, A Dalmia, LM Häsler, M Lambert, M De Vleesch, HA Davies, J Madine, D Kronenberg, R Feederle, D Del Turco, KPR Nilsson, T Lashley, T Deller, M Gearing, LC Walker, P Heutink, F Rousseau, J Schymkowit, M Jucker, JJ Neher
Nature, 2022-11-16;0(0):.
Species: Human
Sample Types: Tissue Homogenates
-
Maternal breast milk microbiota and immune markers in relation to subsequent development of celiac disease in offspring
Authors: J Štšepetova, K Simre, A Tagoma, O Uibo, A Peet, H Siljander, V Tillmann, M Knip, R Mändar, R Uibo
Scientific Reports, 2022-04-22;12(1):6607.
Species: Human
Sample Types: Breast Milk
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MFG-E8 (LACTADHERIN): a novel marker associated with cerebral amyloid angiopathy
Authors: P Marazuela, M Solé, A Bonaterra-, J Pizarro, J Camacho, E Martínez-S, HB Kuiperij, MM Verbeek, AM de Kort, FHBM Schreuder, CJM Klijn, L Castillo-R, O Pancorbo, D Rodríguez-, F Pujadas, P Delgado, M Hernández-
Acta neuropathologica communications, 2021-09-16;9(1):154.
Species: Human
Sample Types: Serum
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Identification and Validation of Nutrient State-Dependent Serum Protein Mediators of Human CD4+ T Cell Responsiveness
Authors: K Han, K Singh, MJ Rodman, S Hassanzade, Y Baumer, RD Huffstutle, J Chen, J Candia, F Cheung, KER Stagliano, M Pirooznia, TM Powell-Wil, MN Sack
Nutrients, 2021-04-28;13(5):.
Species: Human
Sample Types: Serum
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Relationship Between Milk Fat Globule-Epidermal Growth Factor 8 and Intestinal Cytokines in Infants Born Preterm
Authors: JA Asaro, Z Khan, M Brewer, K Klose, C Pesce, RJ Schanler, CN Codipilly
J Pediatr, 2020-11-09;0(0):.
Species: Human
Sample Types: Breast Milk
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Polymorphisms of MFGE8 are associated with susceptibility and clinical manifestations through gene expression modulation in Koreans with systemic lupus erythematosus
Authors: WY Baek, JM Woo, HA Kim, JY Jung, CH Suh
Sci Rep, 2019-12-06;9(1):18565.
Species: Human
Sample Types: Serum
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Functional Assessment of Patient-Derived Retinal Pigment Epithelial Cells Edited by CRISPR/Cas9
Authors: LP Foltz, SE Howden, JA Thomson, DO Clegg
Int J Mol Sci, 2018-12-19;19(12):.
Species: Human
Sample Types: Cell Culture Supernates
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Quantified F-Actin Morphology Is Predictive of Phagocytic Capacity of Stem Cell-Derived Retinal Pigment Epithelium
Authors: C Müller, C Charniga, S Temple, SC Finnemann
Stem Cell Reports, 2018-02-15;0(0):.
Species: Human
Sample Types: Cell Culture Supernates
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Elevated serum MFG-E8 level is possibly associated with the presence of high-intensity cerebral lesions on magnetic resonance imaging in patients with systemic lupus erythematosus
Authors: C Kishi, SI Motegi, O Ishikawa
J. Dermatol, 2017-03-07;0(0):.
Species: Human
Sample Types: Serum
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Correction of MFG-E8 Resolves Inflammation and Promotes Cutaneous Wound Healing in Diabetes
J Immunol, 2016-05-18;0(0):.
Species: Human
Sample Types: Whole Tissue
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Exosomes in Human Breast Milk Promote EMT
Authors: W Qin, Y Tsukasaki, S Dasgupta, N Mukhopadhy, M Ikebe, ER Sauter
Clin Cancer Res, 2016-04-08;0(0):.
Species: Human
Sample Types: Cell Lysates
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MFG-E8 regulates angiogenesis in cutaneous wound healing.
Authors: Uchiyama A, Yamada K, Ogino S, Yokoyama Y, Takeuchi Y, Udey M, Ishikawa O, Motegi S
Am J Pathol, 2014-05-15;184(7):1981-90.
Species: Human
Sample Types: Cell Culture Supernates
FAQs
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Average Rating: 4 (Based on 1 Review)
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Human MFG-E8 Quantikine ELISA Kit
By Adam Guess
on 02/04/2017
Sample Tested: Cell Culture Supernates
