Summary
Precision
Recovery
Linearity
Data Examples
Product Datasheets
Preparation and Storage
Background
Related Research Areas
Mouse 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 (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 (pg/mL) | 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.
Scientific Data
Product Datasheets
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: 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 50 µL of the appropriate Assay Diluent to each well.
- 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.
- Aspirate each well and wash, repeating the process 4 times for a total of 5 washes.
- 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.
- Aspirate and wash 5 times.
- Add 100 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
- 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.
50 µL Assay Diluent
50 µL Standard, Control, or Sample
100 µL Conjugate
100 µL Substrate Solution
100 µL Stop Solution
Citations for Mouse 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.
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Citations: Showing 1 - 8
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Development of human cell-expressed tag-free rhMFG-E8 as a radiation mitigator and a therapeutic for acute kidney injury
Authors: Chaung, W;Zhang, F;Ma, G;Yen, H;Jacob, A;Brenner, M;Wang, P;
Research square
Species: Rat
Sample Types:
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Small extracellular vesicles from Ptpn1-deficient macrophages alleviate intestinal inflammation by reprogramming macrophage polarization via lactadherin enrichment
Authors: D Han, D Lu, S Huang, J Pang, Y Wu, J Hu, X Zhang, Y Pi, G Zhang, J Wang
Redox Biology, 2022-11-28;58(0):102558.
Species: Mouse, Transgenic Mouse
Sample Types: Exosomes
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Increased MFG-E8 at neuromuscular junctions is an exacerbating factor for sarcopenia-associated denervation
Authors: M Ikemoto-Ue, H Zhou, T Kurosawa, Y Yoshimoto, M Toyoda, N Kanazawa, T Nakazawa, M Morita, K Tsuchida, A Uezumi
Aging Cell, 2021-12-24;0(0):e13536.
Species: Mouse
Sample Types: Tissue Homogenates
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MFG-E8 regulated by miR-99b-5p protects against osteoarthritis by targeting chondrocyte senescence and macrophage reprogramming via the NF-&kappaB pathway
Authors: Y Lu, L Liu, J Pan, B Luo, H Zeng, Y Shao, H Zhang, H Guan, D Guo, C Zeng, R Zhang, X Bai, H Zhang, D Cai
Cell Death & Disease, 2021-05-25;12(6):533.
Species: Mouse
Sample Types: Serum
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MFG-E8 Plays an Important Role in Attenuating Cerulein-Induced Acute Pancreatitis in Mice
Authors: HF Bu, S Subramania, H Geng, X Wang, F Liu, PM Chou, C Du, IG De Plaen, XD Tan
Cells, 2021-03-25;10(4):.
Species: Mouse
Sample Types: Plasma
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Living Neurons with Tau Filaments Aberrantly Expose Phosphatidylserine and Are Phagocytosed by Microglia
Authors: J Brelstaff, AM Tolkovsky, B Ghetti, M Goedert, MG Spillantin
Cell Rep, 2018-08-21;24(8):1939-1948.e4.
Species: Mouse
Sample Types: Cell Culture Supernates
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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: Mouse
Sample Types: Cell Culture Supernates
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The neuroprotective effects of milk fat globule-EGF factor 8 against oligomeric amyloid beta toxicity.
J Neuroinflammation, 2012-06-28;9(0):148.
Species: Mouse
Sample Types: Cell Culture Supernates
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