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AB62899

Human Interferon Gamma ELISPOT Kit

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(1 Publication)

Human Interferon Gamma ELISPOT Kit is a highly sensitive immunoassay designed to detect and quantify human IFN-gamma secretion at the single-cell level.

- Ready-to use kits with pre-coated PDVF plates
- Detects individual IFN-Gamma-secreting cells
- Available in single plate or 5 plates format

View Alternative Names

Interferon gamma, IFN-gamma, Immune interferon, IFNG

Key facts

Detection method

Colorimetric

Sample types

Suspension cells

Reacts with

Human

Assay type

Sandwich (qualitative)

Assay Platform

Microplate

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Reactivity data

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

Human Interferon Gamma ELISPOT Kit ab62899 is a sensitive assay for the qualitative measurement of human IFN gamma production and secretion in a single cell suspension

How the assay works

The ELISPOT assay involves a capture antibody highly specific for the analyte of interest coated to the wells of a PVDF bottomed 96 well microtitre plate, either during kit manufacture or in the laboratory. The plate is then blocked to minimize any non-antibody dependent unspecific binding and washed. Cell suspension and stimulant are added and the plate incubated allowing the specific antibodies to bind any analytes produced. Cells are then removed by washing prior to the addition of Biotinylated detection antibodies which bind to the previously captured analyte. Enzyme conjugated streptavidin is then added binding to the detection antibodies. Following incubation and washing substrate is then applied to the wells resulting in colored spots which can be quantified using appropriate analysis software or manually using a microscope.

Human Interferon Gamma ELISPOT Kit ab62899 protocol summary:

1. Add sample (Cells) to appropriate wells. Incubate at 37 C.
2. Aspirate and wash each well. Add prepared Biotinylated labeled detector antibody. Incubate at room temperature.
3. Add prepared Streptavidin-Alkaline Phosphatase mix to each well. Incubate at room temperature.
4. Add the substrate solution BCIP/NBT to each well and monitor spot formation.
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What's included?

{ "values": { "5x96Tests": { "sellingSize": "5 x 96 Tests", "publicAssetCode":"ab62899-5x96Tests", "assetComponentDetails": [ { "size":"5 x 1 Unit", "name":"IFNγ Pre-coated 96-well PDVF-bottomed plates", "number":"AB62899-CMP05" }, { "size":"1 x 50 µL", "name":"Streptavidin - Alkaline Phosphatase conjugated", "number":"AB62899-CMP03" }, { "size":"2 x 25 mL", "name":"Ready-to-use BCIP/NBT substrate buffer", "number":"AB62899-CMP04" }, { "size":"1 x 1 Vial", "name":"Biotinylated Human IFNγ Detection Antibody", "number":"AB62899-CMP01" }, { "size":"1 x 2 g", "name":"Bovine Serum Albumin", "number":"AB62899-CMP02" } ] }, "1x96Tests": { "sellingSize": "1 x 96 Tests", "publicAssetCode":"ab62899-1x96Tests", "assetComponentDetails": [ { "size":"1 x 1 Unit", "name":"IFNγ Pre-coated 96-well PDVF-bottomed plates", "number":"AB62899-CMP05", "productcode":"" }, { "size":"1 x 10 µL", "name":"Streptavidin - Alkaline Phosphatase conjugated", "number":"AB62899-CMP03", "productcode":"" }, { "size":"1 x 2 g", "name":"Bovine Serum Albumin", "number":"AB62899-CMP02", "productcode":"" }, { "size":"1 x 1 Vial", "name":"Biotinylated Human IFNγ Detection Antibody", "number":"AB62899-CMP01", "productcode":"" }, { "size":"1 x 11 mL", "name":"Ready-to-use BCIP/NBT substrate buffer", "number":"AB62899-CMP04", "productcode":"" } ] } } }
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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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Publications (1)

Recent publications for all applications. Explore the full list and refine your search

Cell discovery 10:78 PubMed39039072

2024

Proteogenomic insights into the biology and treatment of pan-melanoma.

Applications

Unspecified application

Species

Unspecified reactive species

Hang Xiang,Rongkui Luo,Yunzhi Wang,Bing Yang,Sha Xu,Wen Huang,Shaoshuai Tang,Rundong Fang,Lingli Chen,Na Zhu,Zixiang Yu,Sujie Akesu,Chuanyuan Wei,Chen Xu,Yuhong Zhou,Jianying Gu,Jianyuan Zhao,Yingyong Hou,Chen Ding
View all publications
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