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AB229415

Human IFN gamma ELISA Kit, Fluorescent

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Human IFN gamma ELISA Kit, Fluorescent is a single-wash 90-min Simplestep used to quantify Human IFN gamma with a sensitivity of 25 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.

- Fluorescent Sandwich ELISA - 530/570/590 nm readout : works on any standard plate reader

View Alternative Names

Interferon gamma, IFN-gamma, Immune interferon, IFNG

3 Images
Sandwich ELISA - Human IFN gamma ELISA Kit, Fluorescent (AB229415)
  • sELISA

Unknown

Sandwich ELISA - Human IFN gamma ELISA Kit, Fluorescent (AB229415)

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, Fluorescent (AB229415)
  • sELISA

Unknown

Sandwich ELISA - Human IFN gamma ELISA Kit, Fluorescent (AB229415)

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.

Sandwich ELISA - Human IFN gamma ELISA Kit, Fluorescent (AB229415)
  • sELISA

Unknown

Sandwich ELISA - Human IFN gamma ELISA Kit, Fluorescent (AB229415)

Example of human Interferon-gamma (IFNG) standard curve in Sample Diluent NS.

The Interferon-gamma (IFNG) standard curve was prepared as described in Section 10.

Key facts

Detection method

Fluorescent

Sample types

Heparin Plasma, Citrate plasma, Cell culture supernatant, Serum, EDTA Plasma

Reacts with

Human

Assay type

Sandwich

Results type

Quantitative

Sensitivity

= 25 pg/mL

Range

0.06 - 30 ng/mL

Assay time

1h 30m

Assay Platform

Pre-coated microplate (12 x 8 well strips)

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Reactivity data

{ "title": "Reactivity Data", "filters": { "stats": ["", "Reactivity", "Dilution Info", "Notes"] }, "values": { "sELISA": { "reactivity":"TESTED_AND_REACTS", "dilution-info":"", "notes":"<p></p>" } } }
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Product details

Interferon-gamma (IFNG) in vitro CatchPoint SimpleStep ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for the quantitative measurement of Interferon-gamma (IFNG) protein in human serum, plasma, and cell culture supernatant samples.

This CatchPoint SimpleStep ELISA kit has been optimized for Molecular Devices Microplate Readers. Click here for a list of recommended Microplate Readers.
If using a Molecular Devices' plate reader supported by SoftMax® Pro software, a preconfigured protocol for these CatchPoint SimpleStep ELISA Kits is available with all the protocol and analysis settings at www.softmaxpro.org.

The CatchPoint SimpleStep ELISA employs an affinity tag labeled capture antibody and a reporter conjugated detector antibody which immunocapture the sample analyte in solution. This entire complex (capture antibody/analyte/detector antibody) is in turn immobilized via immunoaffinity of an anti-tag antibody coating the well. To perform the assay, samples or standards are added to the wells, followed by the antibody mix. After incubation, the wells are washed to remove unbound material. CatchPoint HRP Development Solution containing the Stoplight Red Substrate is added. During incubation, the substrate is catalyzed by HRP generating a fluorescent product. Signal is generated proportionally to the amount of bound analyte and the intensity is measured in a fluorescence plater reader at 530/570/590 nm Excitation/Cutoff/Emission.

IFN gamma 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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Precision

[ { "reproducibilityType": "Inter", "sample": "Supernatant", "replicates": 3, "mean": null, "standardDeviation": null, "coefficientOfVariability": "7.9" }, { "reproducibilityType": "Intra", "sample": "Supernatant", "replicates": 5, "mean": null, "standardDeviation": null, "coefficientOfVariability": "1.1" } ]
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Recovery

[ { "sample": "Cell culture supernatant", "range": "77 - 93 %", "average": "= 86" }, { "sample": "Serum", "range": "98 - 104 %", "average": "= 102" }, { "sample": "EDTA Plasma", "range": "117 - 124 %", "average": "= 122" }, { "sample": "Heparin Plasma", "range": "197 - 298 %", "average": "= 244" }, { "sample": "Citrate plasma", "range": "96 - 115 %", "average": "= 107" } ]
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What's included?

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Properties and storage information

Shipped at conditions
Blue Ice
Appropriate short-term storage conditions
+4°C
Appropriate long-term storage conditions
+4°C
Storage information
+4°C
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Supplementary information

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

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.

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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Product protocols

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Target data

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).
See full target information IFNG
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Product promise

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