Human IFN gamma ELISA Kit (ab174443)
Key features and details
- One-wash 90 minute protocol
- Sensitivity: 470 pg/ml
- Range: 0.468 ng/ml - 30 ng/ml
- Sample type: Cell culture supernatant, Plasma, Serum
- Detection method: Colorimetric
- Assay type: Sandwich (quantitative)
- Reacts with: Human
Overview
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Product name
Human IFN gamma ELISA Kit
See all Interferon gamma kits -
Detection method
Colorimetric -
Precision
Intra-assay Sample n Mean SD CV% Overall 5 1.1% Inter-assay Sample n Mean SD CV% Overall 3 7.9% -
Sample type
Cell culture supernatant, Serum, Plasma -
Assay type
Sandwich (quantitative) -
Sensitivity
470 pg/ml -
Range
0.468 ng/ml - 30 ng/ml -
Recovery
Sample specific recovery Sample type Average % Range Serum 102 98% - 104% Cell culture media 86 77% - 93% Hep Plasma 244 197% - 298% EDTA Plasma 122 117% - 124% Cit plasma 107 96% - 115% -
Assay time
1h 30m -
Assay duration
One step assay -
Species reactivity
Reacts with: Human -
Product overview
Human IFN gamma ELISA kit (ab174443) is a single-wash 90 min sandwich ELISA designed for the quantitative measurement of IFN-gamma protein in human cell culture supernatant, plasma and serum samples. It uses our proprietary SimpleStep ELISA® technology. Quantitate human IFN gamma with 470 pg/ml sensitivity.
SimpleStep ELISA® technology employs capture antibodies conjugated to an affinity tag that is recognized by the monoclonal antibody used to coat our SimpleStep ELISA® plates. This approach to sandwich ELISA allows the formation of the antibody-analyte sandwich complex in a single step, significantly reducing assay time. See the SimpleStep ELISA® protocol summary in the image section for further details. Our SimpleStep ELISA® technology provides several benefits:
-Single-wash protocol reduces assay time to 90 minutes or less
-High sensitivity, specificity and reproducibility from superior antibodies
-Fully validated in biological samples
-96-wells plate breakable into 12 x 8 wells strips
A 384-well SimpleStep ELISA® microplate (ab203359) is available to use as an alternative to the 96-well microplate provided with SimpeStep ELISA® kits.ASSAY SPECIFICITY
This kit recognizes both native and recombinant human Interferon-gamma protein in serum, plasma, and cell culture supernatant samples only.
Urine, milk, and cell and tissue extract samples have not been tested with this kit.
ASSAY INTERFERENCE
This kit is incompatible with plasma (heparin) samples.
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Notes
IFN gamma (IFNG) is produced by lymphocytes activated by specific antigens or mitogens. IFN gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons.
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Platform
Microplate
Properties
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Storage instructions
Store at +4°C. Please refer to protocols. -
Components 1 x 96 tests 10X Human IFNG Capture Antibody 1 x 600µl 10X Human IFNG Detector Antibody 1 x 600µl 10X Wash Buffer PT (ab206977) 1 x 20ml Antibody Diluent CPI - HAMA Blocker (ab193969) 1 x 6ml Human IFNG Lyophilized Recombinant Protein 2 vials Plate Seals 1 unit Sample Diluent NBP 1 x 20ml Sample Diluent NS (ab193972) 1 x 12ml SimpleStep Pre-Coated 96-Well Microplate (ab206978) 1 unit Stop Solution 1 x 12ml TMB Development Solution 1 x 12ml -
Research areas
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Function
Produced by lymphocytes activated by specific antigens or mitogens. IFN-gamma, in addition to having antiviral activity, has important immunoregulatory functions. It is a potent activator of macrophages, it has antiproliferative effects on transformed cells and it can potentiate the antiviral and antitumor effects of the type I interferons. -
Tissue specificity
Released primarily from activated T lymphocytes. -
Involvement in disease
In Caucasians, genetic variation in IFNG is associated with the risk of aplastic anemia (AA) [MIM:609135]. AA is a rare disease in which the reduction of the circulating blood cells results from damage to the stem cell pool in bone marrow. In most patients, the stem cell lesion is caused by an autoimmune attack. T-lymphocytes, activated by an endogenous or exogenous, and most often unknown antigenic stimulus, secrete cytokines, including IFN-gamma, which would in turn be able to suppress hematopoiesis. -
Sequence similarities
Belongs to the type II (or gamma) interferon family. -
Post-translational
modificationsProteolytic processing produces C-terminal heterogeneity, with proteins ending alternatively at Gly-150, Met-157 or Gly-161. -
Cellular localization
Secreted. - Information by UniProt
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Alternative names
- IF 1
- IFG
- IFI
see all -
Database links
- Entrez Gene: 3458 Human
- Omim: 147570 Human
- SwissProt: P01579 Human
- Unigene: 856 Human
Images
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Other - Human IFN gamma ELISA Kit (ab174443)
SimpleStep ELISA technology allows the formation of the antibody-antigen complex in one single step, reducing assay time to 90 minutes. Add samples or standards and antibody mix to wells all at once, incubate, wash, and add your final substrate. See protocol for a detailed step-by-step guide.
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Example IFNG standard curve for cell culture supernatant samples measurements.Background-subtracted data values (mean +/- SD) are graphed. -
Example of human Interferon-gamma standard curve in Sample Diluent NS (for supernatant samples)The Interferon-gamma standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Example IFNG standard curve for serum/plasma samples measurements.Background-subtracted data values (mean +/- SD) are graphed. -
Example of human Interferon-gamma standard curve in Sample Diluent NBP (for serum/plasma samples)The Interferon-gamma standard curve was prepared as described. Raw data values are shown in the table. Background-subtracted data values (mean +/- SD) are graphed.
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Comparison of secreted IFNG in unstimulated and PHA-stimulated Human PBMC.PBMC were grown in the absence or presence of phytohemagglutinin (PHA) for 2 days. IFNG concentrations were measured in 12X and 6X diluted cell culture supernatants of the unstimulated PBMC and the stimulated PBMC, and media. Raw data values (mean +/-SD, n=3) are graphed. The dotted line represents zero sample background. -
Interpolated concentrations of secreted IFNG in unstimulated and PHA-stimulated Human PBMC.The concentrations of IFNG were interpolated from data values shown in Figure 3 using IFNG standard curve and corrected for sample dilution. The mean IFNG concentration was determined to be 1.8 ng/mL in unstimulated PBMC supernatants and 177.2 ng/mL in stimulated PBMC supernatants. -
Linearity of dilution.Linearity of dilution is determined based on interpolated values from the standard curve. Linearity of dilution defines a sample concentration interval in which interpolated target concentrations are directly proportional to sample dilution.
Native Interferon-gamma was measured in PHA-stimulated PBMC cell culture supernatant samples in a 2-fold dilution series. Sample dilutions are made in Sample Diluent NS.
Recombinant Interferon-gamma was spiked into human serum and plasma (citrate and EDTA) samples and diluted in a 2-fold dilution series in Sample Diluent NBP.Neat pooled serum and plasma (EDTA and Citrate) samples from healthy donors was measured in duplicate. All values were below the detectable range of the assay.
Neat serum from ten individual healthy human female/male donors was measured in duplicate. All values were below the detectable range of the assay.
