GM-CSF (Granulocyte-Macrophage Colony Stimulating Factor) induces the development of monocytes, neutrophils, eosinophils, and myeloid and dermal dendritic cells. It also acts as a neutrophil and dendritic cell chemoattractant. GM-CSF promotes Th1 and Th17 cell mediated autoimmune inflammation as well as the inflammatory activation of dendritic cells, microglia, alveolar macrophages, and eosinophils. In addition, it cooperates with G-CSF in promoting tumor cell proliferation and invasion. GM-CSF signals through a receptor complex composed of GM-CSF R alpha and the Common beta Chain (beta c) with Syndecan-2 as a potential co-receptor. The beta c subunit is shared by the receptor complexes for IL-3 and IL-5.
Human GM-CSF Quantikine ELISA Kit
R&D Systems | Catalog # DGM00
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Key Product Details
Assay Length
3.5 hours or 4.5 hours
Sample Type & Volume Required Per Well
Cell Culture Supernates (100 µL), Serum (100 µL), EDTA Plasma (100 µL), Heparin Plasma (100 µL), Citrate Plasma (100 µL)
Sensitivity
3 pg/mL
Assay Range
7.8-500 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma, Citrate Plasma)
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Product Summary for Human GM-CSF Quantikine ELISA Kit
The Quantikine Human GM-CSF Immunoassay is a 3.5 or 4.5 hour solid phase ELISA designed to measure GM-CSF levels in cell culture supernates, serum, and plasma. It contains E. coli-expressed recombinant human GM-CSF and antibodies raised against the recombinant factor. It has been shown to quantitate recombinant human GM-CSF accurately. Results obtained using natural human GM-CSF 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 GM-CSF.
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 GM-CSF
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 - Forty serum and plasma samples from apparently healthy volunteers were
evaluated for the presence of human GM-CSF in this assay. All samples measured less than the
lowest Human GM-CSF Standard, 7.8 pg/mL. No medical histories were available for the donors
used in this study.
Cell Culture Supernates - Human peripheral blood mononuclear cells (1 x 106
cells/mL) were
cultured in RPMI supplemented with 10% fetal bovine serum, 50 μM beta -mercaptoethanol,
2 mM L-glutamine, 100 U/mL penicillin, and 100 μg/mL streptomycin sulfate and cultured
unstimulated or stimulated with 10 μg/mL PHA. Aliquots of the cell culture supernates were
removed on days 1 and 5 and assayed for levels of human GM-CSF.
| Condition | Day 1 (pg/mL) | Day 5 (pg/mL) |
| Unstimulated | 39 | 43 |
| Stimulated | 71 | 31 |
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
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 46.8 | 131 | 269 | 45.1 | 133 | 266 |
| Standard Deviation | 1.1 | 3.6 | 7.3 | 2.4 | 7.0 | 11.5 |
| CV% | 2.4 | 2.7 | 2.7 | 5.3 | 5.3 | 4.3 |
Citrate Plasma, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||||
|---|---|---|---|---|---|---|
| Sample | 1 | 2 | 3 | 1 | 2 | 3 |
| n | 20 | 20 | 20 | 40 | 40 | 40 |
| Mean (pg/mL) | 54.7 | 156 | 321 | 53.1 | 157 | 317 |
| Standard Deviation | 1.1 | 3.2 | 4.8 | 3.3 | 9.1 | 15.4 |
| CV% | 2.0 | 2.1 | 1.5 | 6.2 | 5.8 | 4.9 |
Recovery for Human GM-CSF Quantikine ELISA Kit
The recovery of GM-CSF spiked to three different levels throughout the range of the assay in various matrices was evaluated.
| Sample Type | Average % Recovery | Range % |
|---|---|---|
| Cell Culture Media (n=5) | 97 | 85-104 |
| Citrate Plasma (n=5) | 101 | 92-106 |
| EDTA Plasma (n=5) | 100 | 94-105 |
| Heparin Plasma (n=5) | 98 | 92-104 |
| Serum (n=5) | 98 | 93-103 |
Linearity
To assess the linearity of the assay, five samples were spiked with high concentrations of GM-CSF in various matrices and diluted with the appropriate Calibrator Diluent to produce values within the dynamic range of the assay.
Scientific Data Images for Human GM-CSF Quantikine ELISA Kit
Human GM-CSF ELISA Cell Culture Supernate Standard Curve
Human GM-CSF ELISA Serum/Plasma 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: GM-CSF
Long Name
Granulocyte Macrophage Growth Factor
Alternate Names
CSF-2, CSF2, GMCSF, Molgramostim, Sargramostim
Entrez Gene IDs
Gene Symbol
CSF2
Additional GM-CSF Products
Product Documents for Human GM-CSF 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 GM-CSF 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 Human GM-CSF Quantikine ELISA Kit
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Sample Tested: SerumVerified Customer | Posted 04/05/2022Clear instructions and robust method
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Sample Tested: SerumVerified Customer | Posted 04/24/2017The Human GM-CSF Quantikine ELISA kit is used for diagnostic use.
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Protocols
View specific protocols for Human GM-CSF Quantikine ELISA Kit (DGM00):
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 100 µL of Assay Diluent to each well.
- Add 100 µL of Standard, control, or sample to each well. Cover with a plate sealer, and incubate at room temperature for 2 hours.
- Aspirate each well and wash, repeating the process 3 times for a total of 4 washes.
- Add 200 µL of Conjugate to each well.
- For Serum & Plasma Samples: Cover with a new plate sealer, and incubate at room temperature for 2 hours.
For Cell Culture Supernate Samples: Cover with a new plate sealer, and incubate at room temperature for 1 hour. - Aspirate and wash 4 times.
- Add 200 µL Substrate Solution to each well. Incubate at room temperature for 20 minutes. PROTECT FROM LIGHT.
- 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.
100 µL Assay Diluent
100 µL Standard, Control, or Sample
200 µL Conjugate
200 µL Substrate Solution
50 µL Stop Solution
Find general support by application which include: protocols, troubleshooting, illustrated assays, videos and webinars.
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