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AB64029

Mouse Interferon gamma ELISPOT Kit

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(9 Publications)

Mouse Interferon gamma ELISPOT Kit is a highly sensitive immunoassay designed to detect and quantify 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, Ifng

Key facts

Detection method

Colorimetric

Sample types

Suspension cells

Reacts with

Mouse

Assay type

Sandwich

Results type

Qualitative

Assay Platform

Microplate

Reactivity data

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

Product details

Mouse Interferon gamma ELISPOT Kit ab64029 is a sensitive assay for the qualitative measurement of mouse 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.

Mouse Interferon gamma ELISPOT Kit ab64029 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.

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

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.

Product protocols

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 : 11585387, PubMed : 8456301). Primarily signals through the JAK-STAT pathway after interaction with its receptor IFNGR1 to affect gene regulation. 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. Plays a role in class I antigen presentation pathway by inducing a replacement of catalytic proteasome subunits with immunoproteasome subunits. In turn, increases the quantity, quality, and repertoire of peptides for class I MHC loading. 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. 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 (By similarity). Participates in the regulation of hematopoietic stem cells during development and under homeostatic conditions by affecting their development, quiescence, and differentiation (PubMed : 20535209, PubMed : 25078851).
See full target information Ifng

Publications (9)

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

Nature communications 16:2971 PubMed40140411

2025

An mRNA vaccine against monkeypox virus inhibits infection by co-activation of humoral and cellular immune responses.

Applications

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Species

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Wanbo Tai,Chongyu Tian,Huicheng Shi,Benjie Chai,Xinyang Yu,Xinyu Zhuang,Pengyuan Dong,Min Li,Qi Yin,Shengyong Feng,Weixiao Wang,Oujia Zhang,Shibo Liang,Yang Liu,Jianying Liu,Longchao Zhu,Guangyu Zhao,Mingyao Tian,Guocan Yu,Gong Cheng

Molecular therapy. Nucleic acids 34:102045 PubMed37876532

2023

Reducing cell intrinsic immunity to mRNA vaccine alters adaptive immune responses in mice.

Applications

Unspecified application

Species

Unspecified reactive species

Ziyin Wang,Egon J Jacobus,David C Stirling,Stefanie Krumm,Katie E Flight,Robert F Cunliffe,Jonathan Mottl,Charanjit Singh,Lucy G Mosscrop,Leticia Aragão Santiago,Annette B Vogel,Katalin Kariko,Ugur Sahin,Stephanie Erbar,John S Tregoning

Nature communications 14:2962 PubMed37221158

2023

An mRNA-based T-cell-inducing antigen strengthens COVID-19 vaccine against SARS-CoV-2 variants.

Applications

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Species

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Wanbo Tai,Shengyong Feng,Benjie Chai,Shuaiyao Lu,Guangyu Zhao,Dong Chen,Wenhai Yu,Liting Ren,Huicheng Shi,Jing Lu,Zhuming Cai,Mujia Pang,Xu Tan,Penghua Wang,Jinzhong Lin,Qiangming Sun,Xiaozhong Peng,Gong Cheng

Journal for immunotherapy of cancer 11: PubMed36958764

2023

Adaptation of transgene mRNA translation boosts the anticancer efficacy of oncolytic HSV1.

Applications

Unspecified application

Species

Unspecified reactive species

Huy-Dung Hoang,Aida Said,Nasana Vaidya,Victoria H Gilchrist,Kyle Malone,Usha Kabilan,Serena Topshee,Xiao Xiang,An-Dao Yang,David Olagnier,Karen Mossman,Shawn T Beug,Seyed Mehdi Jafarnejad,Samuel T Workenhe,Tyson E Graber,Tommy Alain

PLoS pathogens 19:e1011090 PubMed36634130

2023

Exosomes mediate Coxsackievirus B3 transmission and expand the viral tropism.

Applications

Unspecified application

Species

Unspecified reactive species

Yuxuan Fu,Sidong Xiong

Molecular therapy. Nucleic acids 31:29-42 PubMed36589712

2023

Formulation, inflammation, and RNA sensing impact the immunogenicity of self-amplifying RNA vaccines.

Applications

Unspecified application

Species

Unspecified reactive species

John S Tregoning,David C Stirling,Ziyin Wang,Katie E Flight,Jonathan C Brown,Anna K Blakney,Paul F McKay,Robert F Cunliffe,Valarmathy Murugaiah,Christopher B Fox,Mitchell Beattie,Ying K Tam,Cecilia Johansson,Robin J Shattock

Proceedings of the National Academy of Sciences of the United States of America 119:e2207841119 PubMed35969778

2022

Lipid nanoparticle-mediated lymph node-targeting delivery of mRNA cancer vaccine elicits robust CD8 T cell response.

Applications

Unspecified application

Species

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Jinjin Chen,Zhongfeng Ye,Changfeng Huang,Min Qiu,Donghui Song,Yamin Li,Qiaobing Xu

Advanced functional materials 31: PubMed35822179

2021

FAP-Targeted Photodynamic Therapy Mediated by Ferritin Nanoparticles Elicits an Immune Response against Cancer Cells and Cancer Associated Fibroblasts.

Applications

Unspecified application

Species

Unspecified reactive species

Shiyi Zhou,Zipeng Zhen,Amy V Paschall,Lijun Xue,Xueyuan Yang,Anne-Gaelle Bebin-Blackwell,Zhengwei Cao,Weizhong Zhang,Mengzhe Wang,Yong Teng,Gang Zhou,Zibo Li,Fikri Y Avci,Wei Tang,Jin Xie

Cell 183:1117-1133.e19 PubMed33096019

2020

Detecting Tumor Antigen-Specific T Cells via Interaction-Dependent Fucosyl-Biotinylation.

Applications

Unspecified application

Species

Unspecified reactive species

Zilei Liu,Jie P Li,Mingkuan Chen,Mengyao Wu,Yujie Shi,Wei Li,John R Teijaro,Peng Wu
View all publications
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