Human Interferon gamma ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Interferon gamma with a sensitivity of 3.428 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 384-well plate for higher throughput
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
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Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture media, Cell culture supernatant, EDTA Plasma, Heparin Plasma, Saliva, Serum
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 7.813 - 500 pg/mL
- Assay time
- 1h 30m
- Sensitivity
- = 3.428 pg/mL
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Reactivity data
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application sELISA | Reactivity Reacts | Dilution info - | Notes - |
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
Interferon gamma, IFN-gamma, Immune interferon, IFNG
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Human Interferon gamma ELISA Kit is a single-wash 90-min Simplestep used to quantify Human Interferon gamma with a sensitivity of 3.428 pg/ml. The assay uses a simple mix-wash-read protocol with just one incubation and one wash step.
- Colorimetric Sandwich ELISA - 450 nm readout : works on any standard plate reader
- Different formats for different needs: 384-well plate for higher throughput
- Design your own immunoassay: we also offer the conjugation-ready antibody pair
Key facts
- Detection method
- Colorimetric
- Sample types
- Cell culture media, Cell culture supernatant, EDTA Plasma, Heparin Plasma, Saliva, Serum
- Assay type
- Sandwich (quantitative)
- Reactive species
- Human
- Range
- 7.813 - 500 pg/mL
- Assay time
- 1h 30m
- Assay Platform
- Pre-coated microplate (12 x 8 well strips)
- Sensitivity
- = 3.428 pg/mL
Precision
Intra assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Supernatant | n 8 | mean - | SD - | C.V. 7.4 |
Inter assay
| Sample | n | mean | SD | C.V. |
|---|---|---|---|---|
Sample Supernatant | n 3 | mean - | SD - | C.V. 11.38 |
Recovery
Sample specific recovery
| Sample type | Average % | Range |
|---|---|---|
Sample type Cell culture supernatant | Average % = 104 | Range 97 - 115 % |
Sample type Citrate plasma | Average % = 101 | Range 95 - 109 % |
Sample type EDTA Plasma | Average % = 100 | Range 93 - 106 % |
Sample type Heparin Plasma | Average % = 103 | Range 93 - 114 % |
Sample type Serum | Average % = 101 | Range 96 - 104 % |
Storage
- Shipped at conditions
- Blue Ice
- Appropriate short-term storage conditions
- +4°C
- Appropriate long-term storage conditions
- +4°C
- Storage information
- +4°C
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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Sandwich ELISA - Human Interferon gamma ELISA Kit (ab300323)
Example of human Interferon-gamma standard curve in 96-well vs. 384-well plate. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Interferon gamma ELISA Kit (ab300323)
Interpolated concentration of native Interferon-gamma was measured in duplicate at different sample concentrations in 96-well vs. 384-well plates. Undiluted samples are 2.5% stimulated PBMC supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human Interferon gamma ELISA Kit (ab300323)
Example of human Interferon-gamma standard curve in Sample Diluent NS.
Example of human Interferon-gamma standard curve. Background-subtracted data values (mean +/- SD) are graphed.
Sandwich ELISA - Human Interferon gamma ELISA Kit (ab300323)
Interpolated concentrations of human Interferon-gamma in stimulated PBMC cell culture supernatant.
Interpolated concentration of native Interferon-gamma was measured in duplicate at different sample concentrations. Undiluted samples are 2.5% stimulated PBMC cell culture supernatant. The interpolated dilution factor corrected values are plotted (mean +/- SD, n=2). Sample dilutions are made in Sample Diluent NS.
Sandwich ELISA - Human Interferon gamma ELISA Kit (ab300323)
Interpolated concentrations of Human Interferon-gamma in PBMC cell culture supernatants.
Interpolated concentration of native Interferon-gamma was measured in duplicate in human PBMC cell culture supernatants. Sample dilutions are made in Sample Diluent NS. Human PBMCs were cultured for 46 hours in the presence or absence of 1.5% PHA-M. The concentrations of human Interferon gamma were measured in three different dilutions of the supernatant samples in duplicates and interpolated from the Interferon gamma standard curve. The interpolated values are plotted (mean +/- SD, n=2).
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