Human MFG-E8 Quantikine ELISA Kit

  (5 citations)
(1 Review)
    
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Product Details
Assay Procedure
Citations (5)
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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), 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.
Control Available
QC120 , Quantikine Immunoassay Control Set 765 for Human MFG-E8 - Please Inquire
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.
 MFG-E8 [HRP]
Product Datasheets

Certificate of Analysis Lookup

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Note: Certificate of Analysis not available for kit components.

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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
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. 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:

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.

5 Citations: Showing 1 - 5
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Species
Sample Type
  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 Type: Cell Culture Supernates
  2. 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 Type: Serum
  3. Correction of MFG-E8 Resolves Inflammation and Promotes Cutaneous Wound Healing in Diabetes
    J Immunol, 2016;0(0):.
    Species: Human
    Sample Type: Whole Tissue
  4. 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 Type: Cell Lysates
  5. 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 Type: Cell Culture Supernates

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ELISA Controls
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Quantikine Immunoassay Control Set 765 for Human MFG-E8

Ctrl QC120
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Supplemental ELISA Products
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Quantikine ELISA Wash Buffer 1

ELISA WA126 6  
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Average Rating: 4 (Based on 1 review)

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 Human MFG-E8 Quantikine ELISA Kit DFGE80 Array DFGE80
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 Human MFG-E8 Quantikine ELISA Kit DFGE80
: Human MFG-E8 Quantikine ELISA Kit [DFGE80].

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