Mouse MFG-E8 Quantikine ELISA Kit

  (2 citations)     
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Product Details
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 (25 uL), Serum (10 uL), Heparin Plasma (10 uL)
  • Sensitivity
    0.02 ng/mL
  • Assay Range
    0.1 - 8 ng/mL
  • Specificity
    Natural and recombinant mouse 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 Mouse MFG-E8 Immunoassay is a 4.5 hour solid phase ELISA designed to measure mouse MFG-E8 in cell culture supernates, serum, and plasma. It contains NS0-expressed recombinant mouse MFG-E8 and antibodies raised against the recombinant factor. This immunoassay has been shown to quantitate the recombinant factor accurately. Results obtained using natural mouse MFG-E8 showed dose-response curves that were parallel to the standard curves obtained using the recombinant kit standards. These results indicate that this kit can be used to determine relative mass values for natural mouse 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 known concentration were tested in separate assays to assess inter-assay precision
Cell Culture Supernates, Serum, Heparin Plasma
Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 0.506 0.936 3.89 0.509 0.91 3.78
Standard Deviation 0.02 0.042 0.116 0.047 0.071 0.322
CV% 4 4.5 3 9.2 7.8 8.5

Recovery

The recovery of MFG-E8 spiked into various matrices was evaluated.

Sample Type Average % Recovery Range %
Cell Culture Supernates (n=4) 99 90-104
Heparin Plasma (n=4) 94 90-98
Serum (n=4) 101 97-104
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of mouse MFG-E8 were serially diluted with Calibrator Diluent to produce samples with values within the dynamic range of the assay.
 MFG-E8 [HRP]
Product Datasheets

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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. 50 µL Assay Diluent
  4.   Add 50 µL of the appropriate 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 4 times for a total of 5 washes.

  8. 100 µL Conjugate
  9.   Add 100 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 5 times.

  11. 100 µL Substrate Solution
  12.   Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.

  13. 100 µL Stop Solution
  14.   Add 100 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. 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: Mouse
    Sample Type: Cell Culture Supernates
  2. The neuroprotective effects of milk fat globule-EGF factor 8 against oligomeric amyloid beta toxicity.
    J Neuroinflammation, 2012;9(0):148.
    Species: Mouse
    Sample Type: Cell Culture Supernates

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