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Human IFN gamma ELISA Kit ab174443 is a single-wash 90-min SimpleStep ELISA used to quantify Human IFN gamma with a sensitivity of 470 pg/ml. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.

Easy-to-use, rapid, sensitive Human IFN gamma sandwich ELISA Kit:
- Mix, Wash, Read: get results in 90 minutes with SimpleStep ELISA® format
- Colorimetric assay - 450nm readout; works on any plate reader
- Different formats for different needs: 384-well for higher throughput

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Images

ELISA - Human IFN gamma ELISA Kit (AB174443), expandable thumbnail
  • ELISA - Human IFN gamma ELISA Kit (AB174443), expandable thumbnail
  • ELISA - Human IFN gamma ELISA Kit (AB174443), expandable thumbnail
  • ELISA - Human IFN gamma ELISA Kit (AB174443), expandable thumbnail
  • Sandwich ELISA - Human IFN gamma ELISA Kit (AB174443), expandable thumbnail

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Key facts

Detection method

Colorimetric

Sample types

Plasma, Cell culture supernatant, Serum

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

0.468 - 30 ng/mL

Assay time

1h 30m

Sensitivity

= 470 pg/mL

Reactivity data

Application

sELISA

Reactivity

Reacts

Dilution info

-

Notes

-

Associated Products

Select an associated product type

1 product for Alternative Product

Target data

Function

Type II interferon produced by immune cells such as T-cells and NK cells that plays crucial roles in antimicrobial, antiviral, and antitumor responses by activating effector immune cells and enhancing antigen presentation (PubMed:16914093, PubMed:8666937). Primarily signals through the JAK-STAT pathway after interaction with its receptor IFNGR1 to affect gene regulation (PubMed:8349687). Upon IFNG binding, IFNGR1 intracellular domain opens out to allow association of downstream signaling components JAK2, JAK1 and STAT1, leading to STAT1 activation, nuclear translocation and transcription of IFNG-regulated genes. Many of the induced genes are transcription factors such as IRF1 that are able to further drive regulation of a next wave of transcription (PubMed:16914093). Plays a role in class I antigen presentation pathway by inducing a replacement of catalytic proteasome subunits with immunoproteasome subunits (PubMed:8666937). In turn, increases the quantity, quality, and repertoire of peptides for class I MHC loading (PubMed:8163024). Increases the efficiency of peptide generation also by inducing the expression of activator PA28 that associates with the proteasome and alters its proteolytic cleavage preference (PubMed:11112687). Up-regulates as well MHC II complexes on the cell surface by promoting expression of several key molecules such as cathepsins B/CTSB, H/CTSH, and L/CTSL (PubMed:7729559). Participates in the regulation of hematopoietic stem cells during development and under homeostatic conditions by affecting their development, quiescence, and differentiation (By similarity).

Alternative names

What's included?

1 x 384 Tests
Components
10X Human IFNG Capture Antibody
1 x 600 µL
10X Human IFNG Detector Antibody
1 x 600 µL
10X Wash Buffer PT (ab206977)
1 x 20 mL
384 well CaptSure™ microplates
1 x 1 Unit
Antibody Diluent CPI2
1 x 6 mL
Human IFNG Lyophilized Recombinant Protein
2 x 1 Vial
Plate Seals
1 x 1 Unit
Sample Diluent NBP
1 x 20 mL
Sample Diluent NS (ab193972)
1 x 100 mL
Stop Solution
2 x 12 mL
TMB Development Solution
2 x 12 mL

Recommended products

Human IFN gamma ELISA Kit ab174443 is a single-wash 90-min SimpleStep ELISA used to quantify Human IFN gamma with a sensitivity of 470 pg/ml. The assay uses a simple Mix-Wash-Read protocol with just one incubation and wash step.

Easy-to-use, rapid, sensitive Human IFN gamma sandwich ELISA Kit:
- Mix, Wash, Read: get results in 90 minutes with SimpleStep ELISA® format
- Colorimetric assay - 450nm readout; works on any plate reader
- Different formats for different needs: 384-well for higher throughput

Key facts

Detection method

Colorimetric

Sample types

Plasma, Cell culture supernatant, Serum

Assay type

Sandwich (quantitative)

Reactive species

Human

Range

0.468 - 30 ng/mL

Assay time

1h 30m

Assay Platform

Microplate

Sensitivity

= 470 pg/mL

Precision

Intra assay

Sample

Overall

n

5

mean

-

SD

-

C.V.

1.1

Inter assay

Sample

Overall

n

3

mean

-

SD

-

C.V.

7.9

Recovery

Sample specific recovery

Sample type

Serum

Average %

= 102

Range

98 - 104 %

Sample type

Cell culture media

Average %

= 86

Range

77 - 93 %

Sample type

Citrate plasma

Average %

= 107

Range

96 - 115 %

Sample type

EDTA Plasma

Average %

= 122

Range

117 - 124 %

Sample type

Heparin Plasma

Average %

= 244

Range

197 - 298 %

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

+4°C

Storage information

+4°C

Notes

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 (Pre-coated 384 well Microplate SimpleStep ELISA® 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.

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.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Interferon gamma (IFN-γ) also known as type II interferon is a cytokine that plays an important role in immune response. IFN-γ has a molecular weight of about 17 kDa and is produced by T cells and natural killer (NK) cells. IFN-γ binds to the interferon gamma receptor initiating a signaling cascade that activates various genes involved in immune functions. It is expressed mainly in activated immune cells within lymphoid tissues and inflamed sites during immune responses.

Biological function summary

This cytokine is significant in promoting macrophage activation enhancing the antigen presentation process and boosting the antimicrobial activity of phagocytes. IFN-γ is not part of a larger protein complex but works as a homodimer in signal transduction. Its production heightens the Th1 immune response by stimulating the differentiation of naïve T cells into Th1 cells which is essential for effective cellular immunity.

Pathways

IFN-γ is integrally involved in the JAK-STAT signaling pathway alongside another critical cytokine Interleukin-12. This pathway further amplifies the immune response by regulating the expression of genes associated with cellular defense mechanisms. IFN-γ also interacts with the NF-kB pathway influencing inflammation and the activation of further immune responses. These interactions show a network of cooperativity with proteins like STAT1 and NF-kB essential for executing its biological roles.

Associated diseases and disorders

IFN-γ is linked to autoimmune diseases such as rheumatoid arthritis and multiple sclerosis where its elevated levels can exacerbate inflammatory processes. It connects to other proteins like TNF-alpha in promoting the inflammatory cascade. Moreover lower levels of IFN-γ are associated with a heightened risk of infections like tuberculosis demonstrating its vital role in pathogen defense. Therefore understanding IFN-γ and its interactions can be key in developing therapeutic approaches against these conditions.

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10 product images

  • ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    ELISA - Human IFN gamma ELISA Kit (ab174443)

    Example IFNG standard curve for cell culture supernatant samples measurements. Background-subtracted data values (mean +/- SD) are graphed.

  • ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    ELISA - Human IFN gamma ELISA Kit (ab174443)

    Example IFNG standard curve for serum/plasma samples measurements. Background-subtracted data values (mean +/- SD) are graphed.

  • ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    ELISA - Human IFN gamma ELISA Kit (ab174443)

    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.

  • ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    ELISA - Human IFN gamma ELISA Kit (ab174443)

    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.

  • Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443)

    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.

  • Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443)

    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.

  • Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443)

    Interpolated concentration of native IFN-gamma was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 20% treated human PBMC supernatant (5% PHA-M for 46hr). The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.

  • Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443)

    Example of human IFN-gamma standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443)

    Example of human IFN-gamma standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.

  • Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443), expandable thumbnail

    Sandwich ELISA - Human IFN gamma ELISA Kit (ab174443)

    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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