Human MFG-E8 Quantikine ELISA Kit

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DFGE80
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MFG-E8 ELISA Kit ELISA
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Product Details
Procedure
Citations (6)
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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) <div>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. </div>

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 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 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.
Human MFG-E8 ELISA Linearity

Data Examples

Human MFG-E8 ELISA Serum/Plasma/Saliva Standard Curve

Human MFG-E8 ELISA Cell Culture Supernate/Urine/Human Milk Standard Curve

Product Datasheets

Preparation and Storage

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 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. 100 µL Standard, Control, or Sample
  6.   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.
  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 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.

6 Citations: Showing 1 - 6
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  1. 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;0(0):.
    Species: Human
    Sample Types: Cell Culture Supernates
  2. 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;19(12):.
    Species: Human
    Sample Types: Cell Culture Supernates
  3. 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;0(0):.
    Species: Human
    Sample Types: Serum
  4. Correction of MFG-E8 Resolves Inflammation and Promotes Cutaneous Wound Healing in Diabetes
    J Immunol, 2016;0(0):.
    Species: Human
    Sample Types: Whole Tissue
  5. Exosomes in Human Breast Milk Promote EMT
    Authors: W Qin, Y Tsukasaki, S Dasgupta, N Mukhopadhy, M Ikebe, ER Sauter
    Clin Cancer Res, 2016;0(0):.
    Species: Human
    Sample Types: Cell Lysates
  6. 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;184(7):1981-90.
    Species: Human
    Sample Types: Cell Culture Supernates

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Human MFG-E8 Quantikine ELISA Kit
By Anonymous on 02/04/2017
Sample Tested: Cell Culture Supernates