Human IFN-gamma ELISA - Quantikine QuicKit Best Seller
R&D Systems, part of Bio-Techne | Catalog # QK285
Key Product Details
Assay Length
80 minutes
Sample Type & Volume Required Per Well
Cell Culture Supernates (50 µL), Serum (50 µL), EDTA Plasma (50 µL), Heparin Plasma (50 µL)
Sensitivity
2.56 pg/mL
Assay Range
31.3-2000 pg/mL (Cell Culture Supernates, Serum, EDTA Plasma, Heparin Plasma)
Product Summary for Human IFN-gamma Quantikine QuicKit ELISA
The Quantikine QuicKit Human IFN-gamma Immunoassay is a one step, 80-minute solid phase ELISA designed to measure human IFN-gamma levels in cell culture supernates, serum, and plasma. It contains HEK293-expressed recombinant human IFN-gamma and antibodies raised against the recombinant protein. Results obtained using natural human IFN-gamma showed linear curves that were parallel to the standard curves obtained using the QuicKit standards. These results indicate that this kit can be used to determine relative mass values for natural human IFN-gamma.
Product Specifications
Measurement
Quantitative ELISA
Detection Method
Colorimetric - 450nm (TMB)
Conjugate
HRP
Reactivity
Human
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 - Ten serum and plasma samples from apparently healthy volunteers were
evaluated for the presence of human IFN-gamma in this assay. No medical histories were available for
the donors used in this study. All samples measured less than the lowest standard, 31.3 pg/mL.
ND=Non-detectable
Cell Culture Supernates:
Peripheral blood mononuclear cells (multiple donors) (seeded at 1 x 106
/mL) were cultured in
RPMI 1640 supplemented with 10% fetal bovine serum, 2 mM L-glutamine, 100 U/mL penicillin,
and 100 μg/mL streptomycin sulfate. Cells were left untreated or treated with 10 μg/mL of
PHA for 5 days. Aliquots of the cell culture supernates were removed and assayed for levels of
human IFN-gamma.
CD4+ T cells were isolated from human PBMCs (single donor) using the MagCellect™ Human
CD4+ T cell Isolation Kit (R&D Systems, Catalog # MAGH102). CD4+ T cells were then seeded at
5 x 105
/mL and cultured in ExCellerate Human T Cell Expansion Media, Xeno-Free (R&D
Systems, Catalog # CCM030). T cells were left untreated or treated for 5 days. Treated T cell
media was supplemented with 10 ng/mL GMP recombinant human (rh) IL-7 (R&D Systems,
Catalog # 207-GMP) and 10 ng/mL GMP rhIL-15 (R&D Systems, Catalog # 247-GMP). TCR
stimulation was mediated using either (1) immobilized Human CD3 epsilon Antibody (R&D
Systems, Catalog # MAB100, coated at 1 μg/mL) with 5 μg/mL soluble Human CD28 Antibody
(R&D Systems, Catalog # MAB342) or (2) 25 μL Cloudz CD3/28 particles (Cloudz™ T Cell
Activation Kit-CD3/CD28, R&D Systems, Catalog # CLD001) per mL of culture media. Aliquots of
the cell culture supernates were removed and assayed for levels of human IFN-gamma.
| Condition | (pg/mL) |
| Unstimulated | ND |
| Stimulated PBMCs | 111,232 |
| Unstimulated CD4+ T Cell | ND |
| CD4+ T Cell CD3/CD28 | 52,293 |
| CD4+ T Cell Cloudz | 191,034 |
Precision
Intra-Assay Precision (Precision within an assay) Two samples of known concentration were tested twenty times on one plate to assess intra-assay precision.
Inter-Assay Precision (Precision between assays) Two samples of known concentration were tested in ten separate assays to assess inter-assay precision. Assays were performed by at least three technicians.
Cell Culture Supernates, EDTA Plasma, Heparin Plasma, Serum
| Intra-Assay Precision | Inter-Assay Precision | |||
|---|---|---|---|---|
| Sample | 1 | 2 | 1 | 2 |
| n | 20 | 20 | 10 | 10 |
| Mean (pg/mL) | 198 | 1156 | 211 | 1233 |
| Standard Deviation | 3.76 | 20.2 | 20.4 | 56.9 |
| CV% | 1.9 | 1.8 | 9.6 | 4.6 |
Recovery for Human IFN-gamma Quantikine QuicKit ELISA
The recovery of human IFN-gamma 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) | 92 | 83-102 |
| EDTA Plasma (n=4) | 107 | 88-132 |
| Serum (n=4) | 93 | 74-114 |
Linearity
To assess the linearity of the assay, samples containing and/or spiked with high concentrations of human IFN-gamma were diluted with calibrator diluent to produce samples with values within the dynamic range of the assay. Samples were diluted prior to assay.
Scientific Data Images for Human IFN-gamma Quantikine QuicKit ELISA
Human IFN-gamma QuicKit ELISA Spiked Recovery Competitor Comparison
IFN-gamma is spiked at three known concentrations throughout the range of the assay and run to measure response of the spiked sample matrix. Serum recovery is 115% compared to 144% for the top competitor. Plasma recovery is 120% compared to 135% for the top competitor. Cell Culture Supernates recovery is 118% compared to 79% for the top competitor. In spike and recovery experiments, natural samples are spiked with the recombinant target analyte of interest to identify interference caused by sample matrices.
Human IFN-gamma QuicKit ELISA Spiked Linearity Competitor Comparison
IFN-gamma is spiked at high concentration in various matrices and diluted with appropriate Calibrator Diluent to produce samples with values within the dynamic range of the assay. The linearity is between 99%-126% compared to 71%-85% for the top competitor.
Human IFN-gamma 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: IFN-gamma
Long Name
Interferon gamma
Alternate Names
IFG, IFI, IFNG, IFNgamma
Entrez Gene IDs
Gene Symbol
IFNG
Additional IFN-gamma Products
Product Documents for Human IFN-gamma Quantikine QuicKit ELISA
Product Specific Notices for Human IFN-gamma Quantikine QuicKit ELISA
For research use only
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