Human Interferon gamma + IL4 ELISPOT Kit is a ELISPOT Kit for the sensitive detection of Human Interferon gamma + IL4 production in a single cell suspension.
| Application | Reactivity | Dilution info | Notes |
|---|---|---|---|
Application ELISPOT | Reactivity Reacts | Dilution info - | Notes - |
Cytokine secreted primarily by mast cells, T-cells, eosinophils, and basophils that plays a role in regulating antibody production, hematopoiesis and inflammation, and the development of effector T-cell responses (PubMed:1993171, PubMed:3016727). Induces the expression of class II MHC molecules on resting B-cells. Enhances both secretion and cell surface expression of IgE and IgG1 (PubMed:1993171). Regulates also the expression of the low affinity Fc receptor for IgE (CD23) on both lymphocytes and monocytes (PubMed:2521231). Positively regulates IL31RA expression in macrophages. Stimulates autophagy in dendritic cells by interfering with mTORC1 signaling and through the induction of RUFY4. In addition, plays a critical role in higher functions of the normal brain, such as memory and learning (By similarity). Upon binding to IL4, IL4R receptor dimerizes either with the common IL2R gamma chain/IL2RG to produce the type 1 signaling complex, located mainly on hematopoietic cells, or with the IL13RA1 to produce the type 2 complex, which is expressed also on nonhematopoietic cells (PubMed:10219247, PubMed:11526337, PubMed:18243101). Engagement of both types of receptors initiates JAK3 and to a lower extend JAK1 phosphorylation leading to activation of the signal transducer and activator of transcription 6/STAT6 (PubMed:7721895).
IL4, IL4
Interleukin-4, IL-4, B-cell stimulatory factor 1, Binetrakin, Lymphocyte stimulatory factor 1, Pitrakinra, BSF-1, IL4
Human Interferon gamma + IL4 ELISPOT Kit is a ELISPOT Kit for the sensitive detection of Human Interferon gamma + IL4 production in a single cell suspension.
The ELISPOT assay is designed to enumerate cytokine producing cells in a single cell suspension. This method has the advantage of requiring a minimum of in-vitro manipulations allowing cytokine production analysis as close as possible to in-vivo conditions in a highly specific way. This technique is designed to determine the frequency of cytokine producing cells under a given stimulation, and the follow-up of such frequency during a treatment and/or a pathological state. Elispot assay constitutes an ideal tool in the TH1/TH2 response, vaccine development, viral infection monitoring and treatment, cancerology, infectious diseases, autoimmune diseases and transplantation.
This Elispot assay is based on sandwich immuno-enzyme technology. Cell secreted cytokines or soluble molecules are captured by coated antibodies avoiding diffusion in supernatant, protease degradation or binding on soluble membrane receptors. After cell removal, the captured cytokines are revealed by tracer antibodies and appropriate conjugates.
The dual colour Elispot allows you to monitor the production of two cytokines simultaneously in the same well.
Principle of the method
After cell stimulation, locally produced cytokines are captured by IFN gamma and IL4 specific monoclonal antibodies. After cell lysis, trapped cytokine molecules are revealed by a secondary anti-IFN gamma FITC-conjugated antibody and a biotinylated anti-IL4 antibody. Those are in turn recognised by anti-FITC HRP and streptavidin-AP conjugates. PVDF-bottomed-well plates are then incubated first with AEC substrate buffer, washed and subsequently incubated with BCIP/NBT. Coloured red/brownish spots indicate IFN gamma production while IL4 is revealed by blue/purple spots.
Store plates at room temperature.
Interferon gamma (IFN-γ) and Interleukin 4 (IL4) are cytokines with distinct roles in the immune system. Interferon gamma occasionally referred to as IFN-gamma cytokine is a type II interferon with a molecular mass of approximately 20 kDa. It is expressed by activated T cells and natural killer cells. IL4 on the other hand is a cytokine mainly produced by CD4+ T cells mast cells and basophils. IL4 has a molecular weight of about 15 kDa. Both cytokines are active in various tissues including lymphoid organs and sites of inflammation.
The combination of interferon gamma and IL4 influences immune responses and cellular activities. IFN-γ is known for activating macrophages and inhibiting viral replication while IL4 promotes differentiation of naive helper T cells to Th2 cells. IL4 also supports antibody class switching to IgE. These cytokines do not form a complex; instead they modulate immune responses through their respective receptors and associated signaling pathways.
Interferon gamma and IL4 play key roles in the immune response pathways. IFN-γ is a critical part of the JAK-STAT signaling pathway influencing gene expression related to immune response. It is often associated with proteins such as STAT1 in this pathway. IL4 also interacts with the JAK-STAT pathway but it signals through a different receptor complex involving STAT6 leading to promotion of Th2 cell differentiation and B cell maturation.
Interferon gamma and IL4 have connections to autoimmune diseases and allergies. IFN-γ has been linked to autoimmune disorders like multiple sclerosis where its overexpression can lead to tissue damage. In contrast IL4 is associated with allergic conditions such as asthma where it promotes IgE-mediated hypersensitivity. The dysregulation of these cytokines and associated proteins like STAT1 for IFN-γ and STAT6 for IL4 can aggravate these conditions.
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