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Human GM-CSF Quantikine ELISA Kit

R&D Systems, part of Bio-Techne | Catalog # DGM00

R&D Systems, part of Bio-Techne
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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)

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

Reactivity

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.

ConditionDay 1 (pg/mL)Day 5 (pg/mL)
Unstimulated 3943
Stimulated7131

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.

Human GM-CSF ELISA Linearity

Scientific Data Images for Human GM-CSF Quantikine ELISA Kit

Human GM-CSF ELISA Cell Culture Supernate Standard Curve

Human GM-CSF ELISA Cell Culture Supernate Standard Curve

Human GM-CSF ELISA Serum/Plasma 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

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.

Long Name

Granulocyte Macrophage Growth Factor

Alternate Names

CSF-2, CSF2, GMCSF, Molgramostim, Sargramostim

Entrez Gene IDs

1437 (Human); 12981 (Mouse); 116630 (Rat); 397208 (Porcine); 403923 (Canine); 493805 (Feline)

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 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

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⚠ 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.