GDF-15 (Growth Differentiation Factor 15) is a dimeric TGF-beta superfamily cytokine that is highly expressed in placenta and brain. It promotes neuronal survival and regulates neuropeptide Y and pro-opiomelanocortin levels to suppress appetite. It is cardioprotective via inhibition of platelet activation, limiting atherosclerosis, promoting recovery following myocardial infarction, and regulating angiogenesis. GDF-15 also inhibits cartilage formation and the production of TNF-alpha production from LPS-stimulated macrophages.
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Mouse/Rat GDF-15 Quantikine ELISA Kit
R&D Systems | Catalog # MGD150
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Key Product Details
Assay Length
4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL), Urine (10 µL)
Sensitivity
2.2 pg/mL
Assay Range
7.8-500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
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Product Summary for Mouse/Rat GDF-15 Quantikine ELISA Kit
The Quantikine Mouse/Rat GDF-15 Immunoassay is a 4.5 hour solid-phase ELISA designed to measure mouse and rat GDF-15 in cell culture supernates, serum, plasma, and urine. It contains E.
coli-expressed recombinant mouse GDF-15 and antibodies raised against the recombinant factor. This immunoassay has been shown to accurately quantitate the recombinant factor. Results obtained using natural mouse or rat GDF-15 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 naturally occurring mouse and rat GDF-15.
Product Specifications
Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Species
Mouse, Rat
Specificity
Natural mouse and rat GDF-15, recombinant mouse mature GDF-15, precursor GDF-15, and GDF-15/BMP-2 heterodimer. It does not detect monomeric recombinant mouse GDF-15.
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.
Sample Values
Serum/Plasma - Samples were evaluated for the presence of mouse/rat GDF-15 in this assay.
ND=Non-detectable
| Mouse Samples | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=10) | 77.0 | 47.4-97.1 | 19.5 |
| EDTA plasma (n=5) | 241 | 54.4-874 | 354 |
| Heparin plasma (n=5) | 110 | 82.4-169 | 34.7 |
| Urine (n=10) | 2376 | 632-6255 | 1547 |
| Rat Samples | Mean (pg/mL) | Range (pg/mL) | Standard Deviation (pg/mL) |
| Serum (n=10) | 27.5 | 10.5-49.4 | 14.3 |
| Heparin plasma (n=5) | 42.7 | 11.1-145 | 57.3 |
| Urine (n=10) | 4972 | 1922-11,550 | 2747 |
| Rat Samples | Mean of Detectable (pg/mL) | % Detectable | Standard Deviation (pg/mL) |
| EDTA plasma (n=5) | 31.5 | 80 | ND-68.8 |
Cell Culture Supernates:
C2C12 mouse myoblast cells were cultured in DMEM supplemented with 10% fetal bovine
serum to 80% confluency. Cell were washed with PBS and cultured in DMEM supplemented
with 5% equine serum for 6 days with media changes every other day. When the cells were
50% differentiated, they were unstimulated or stimulated with 1000 μM H2O2 for 1 day. Aliquots
of the cell culture supernates were removed, assayed for mouse/rat GDF-15, and measured
4630 pg/mL and 7450 pg/mL, respectively.
Hepa 1-6 mouse hepatoma cells (1 x 106
cells/mL) were cultured for 6 days in DMEM
supplemented with 10% fetal bovine serum and 2 mM L-glutamine. An aliquot of the cell
culture supernate was removed, assayed for mouse/rat GDF-15, and measured 6255 pg/mL.
H4-ll-E-C3 rat hepatoma cells (2 x 106
cells/mL) were cultured for 3 days in DMEM
supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin, and
100 μg/mL of streptomycin sulfate. An aliquot of the cell culture supernate was removed,
assayed for mouse/rat GDF-15, and measured 514 pg/mL.
Precision
Intra-Assay Precision (Precision within an assay) Three samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Three samples of 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 components.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum, Urine
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 20 | 20 | 20 |
| Mean (pg/mL) | 25.6 | 60.0 | 190 | 26.7 | 63.1 | 196 |
| Standard Deviation | 0.690 | 1.42 | 5.49 | 1.62 | 2.84 | 10.7 |
| CV% | 2.7 | 2.4 | 2.9 | 6.1 | 4.5 | 5.5 |
Recovery for Mouse/Rat GDF-15 Quantikine ELISA Kit
The recovery of GDF-15 spiked to levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Mouse Cell Culture Media (n=4) | 102 | 91-110 |
| Mouse EDTA Plasma (n=4) | 95 | 81-109 |
| Mouse Heparin Plasma (n=4) | 95 | 87-104 |
| Mouse Serum (n=4) | 91 | 82-101 |
| Mouse Urine (n=4) | 97 | 88-106 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of GDF-15 in each matrix were diluted with Calibrator Diluent and assayed.
Scientific Data Images for Mouse/Rat GDF-15 Quantikine ELISA Kit
Mouse/Rat GDF-15 ELISA Standard Curve
Preparation and Storage
Shipping
The product is shipped at ambient temperature. 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: GDF-15
Long Name
Growth Differentiation Factor 15
Alternate Names
GDF15, MIC-1, NAG-1, PDF, PLAB, PTGF-beta
Gene Symbol
GDF15
Additional GDF-15 Products
Product Documents for Mouse/Rat GDF-15 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 Mouse/Rat GDF-15 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.Related Research Areas
Citations for Mouse/Rat GDF-15 Quantikine ELISA Kit
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Sample Tested: Mouse plasma and EDTA PlasmaVerified Customer | Posted 07/26/2020
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Protocols
View specific protocols for Mouse/Rat GDF-15 Quantikine ELISA Kit (MGD150):
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.
- 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 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 3 times for a total of 4 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 4 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
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