Human MFG-E8 Quantikine ELISA Kit

R&D Systems | Catalog # DFGE80

R&D Systems
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Key Product Details

Assay Length

4.5 hours

Sample Type & Volume Required Per Well

Cell Culture Supernates (50 µL), Serum (20 µL), EDTA Plasma (20 µL), Heparin Plasma (20 µL), Saliva (50 µL), Urine (50 µL), Human Milk (10 µL)

Sensitivity

13.4 pg/mL

Assay Range

62.5-4000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Saliva, Urine, Human Milk)
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Product Summary for Human MFG-E8 Quantikine ELISA Kit

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.

Product Specifications

Assay Type

Solid Phase Sandwich ELISA

Format

96-well strip plate

Measurement

Quantitative ELISA

Detection Method

Colorimetric - 450nm (TMB)

Conjugate

HRP

Species

Human

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.

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, Human Milk, Urine

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 88.0 125
CV% 4.5 3.3 4.0 8.3 7.0 5.7

EDTA Plasma, Heparin Plasma, Saliva, Serum

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 78.0 117
CV% 6.1 4.1 3.7 7.7 5.9 5.0

Recovery for Human MFG-E8 Quantikine ELISA Kit

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

Scientific Data Images for Human MFG-E8 Quantikine ELISA Kit

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

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

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

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

Preparation and Storage

Shipping

The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below.

Stability & 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

Alternate Names

BA46, Breast epithelial antigen BA46, Lactahedrin, Medin, MFGE8, SED1

Entrez Gene IDs

4240 (Human); 17304 (Mouse)

Gene Symbol

MFGE8

Additional MFG-E8 Products

Product Documents for Human MFG-E8 Quantikine ELISA Kit

Certificate of Analysis

To download a Certificate of Analysis, please enter a lot or batch number in the search box below.

Note: Certificate of Analysis not available for kit components.

Product Specific Notices for Human MFG-E8 Quantikine ELISA Kit

For research use only

⚠ 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.

Citations for Human MFG-E8 Quantikine ELISA Kit

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  • Human MFG-E8 Quantikine ELISA Kit
    Name: Adam Guess
    Sample Tested: Cell Culture Supernates
    Verified Customer | Posted 02/04/2017
    Human MFG-E8 Quantikine ELISA Kit DFGE80

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Protocols

View specific protocols for Human MFG-E8 Quantikine ELISA Kit (DFGE80):

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
  3.   Add 100 µL of Assay Diluent to each well.
    4. 100 µL Standard, Control, or Sample
  4.   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.
  5.   Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
    6. 200 µL Conjugate
  6.   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.
  7.   Aspirate and wash 4 times.
    8. 200 µL Substrate Solution
  8.   Add 200 µL Substrate Solution to each well. Incubate at room temperature for 30 minutes on the benchtop. PROTECT FROM LIGHT.
    9. 50 µL Stop Solution
  9.   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.

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