Mouse/Rat GDF-15 Quantikine ELISA Kit

Catalog # Availability Size / Price Qty
MGD150
Mouse/Rat GDF-15 Standard Curve
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Mouse/Rat GDF-15 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 (50 uL), Serum (50 uL), EDTA Plasma (50 uL), Heparin Plasma (50 uL), Urine (10 uL)
Sensitivity
2.2 pg/mL
Assay Range
7.8 - 500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Urine)
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.

Product Summary

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.

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, Serum, EDTA Plasma, Heparin Plasma, Urine

Intra-Assay Precision Inter-Assay Precision
Sample 1 2 3 1 2 3
n 20 20 20 20 20 20
Mean 25.6 60 190 26.7 63.1 196
Standard Deviation 0.69 1.42 5.49 1.62 2.84 10.7
CV% 2.7 2.4 2.9 6.1 4.5 5.5

Recovery

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.
Mouse/Rat GDF-15 ELISA Linearity

Product Datasheets

Preparation and Storage

Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: GDF-15

Growth/differentiation factors (GDF-1 to GDF-15) are members of the BMP family of TGF-beta superfamily proteins. They are produced as inactive preproproteins which are then cleaved and assembled into active secreted homodimers. GDF dimers are disulfide-linked with the exception of GDF-3 and -9. GDF proteins are important during embryonic development, particularly in the skeletal, nervous, and muscular systems.

Long Name:
Growth Differentiation Factor 15
Entrez Gene IDs:
9518 (Human); 23886 (Mouse); 29455 (Rat)
Alternate Names:
GDF15; GDF-15; growth differentiation factor 15; growth/differentiation factor 15; Macrophage inhibitory cytokine 1; MIC-1; MIC-1NSAID-activated gene 1 protein; MIC1Prostate differentiation factor; NAG-1; NAG-1NSAID-regulated gene 1 protein; NSAID (nonsteroidal inflammatory drug)-activated protein 1; PDF; PDFGDF-15; PLAB; PLABNRG-1; Placental bone morphogenetic protein; Placental TGF-beta; PTGF-beta; PTGFBPTGF-beta
⚠ 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 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 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 3 times for a total of 4 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 4 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.

FAQs

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Mouse/Rat GDF-15 Quantikine ELISA Kit
By Anonymous on 02/02/2017
Sample Tested: cell media