Description: This fc23cpg monoclonal antibody reacts with the p19 subunit of mouse IL-23, a heterodimeric cytokine made up of two disulfide-linked subunits, p40 and p19. It is closely related to IL-12, with which it shares the p40 subunit. IL-23 signals through a heterodimeric receptor expressed on activated T cells, macrophages, and dendritic cells. The receptor shares the IL-12Rbeta1 chain with IL-12, while the IL-23R chain is unique to the IL-23 receptor complex. Binding of IL-23 to its receptor activates Jak/STAT pathways similarly to IL-12, but the two cytokines exhibit distinct functions. Unlike IL-12, which induces T-bet and Th1 differentiation, IL-23 signaling results in RORgammat expression and supports CD4^+ T cells with the Th17 phenotype. Dendritic cells and macrophages produce IL-23 in response to TLR2, TLR4, and TLR8 ligands, as well as agonists of the beta-glucan receptor, Dectin-1. Our studies suggest donor, kinetics and type of stimulant can result in variation in IL-23 expression levels. Recent publications suggest the p19 subunit may also exist in the absence of association with p40. Applications Reported: This fc23cpg antibody has been reported for use in intracellular staining followed by flow cytometric analysis.Applications Tested: This fc23cpg antibody has been tested by intracellular staining and flow cytometric analysis of stimulated mouse bone marrow-derived dendritic cells using the Intracellular Fixation & Permeabilization Buffer Set (cat. 88-8824) and protocol. Please refer to Best Protocols: Protocol A: Two step protocol for (cytoplasmic) intracellular proteins located under the Resources Tab online. This can be used at less than or equal to 0.125 µg per test. A test is defined as the amount (µg) of antibody that will stain a cell sample in a final volume of 100 µL. Cell number should be determined empirically but can range from 10^5 to 10^8 cells/test. It is recommended that the antibody be carefully titrated for optimal performance in the assay of interest. PerCP-eFluor® 710 emits at 710 nm and is excited with the blue laser (488 nm); it can be used in place of PerCP-Cyanine5.5. We recommend using a 710/50 bandpass filter, however, the 695/40 bandpass filter is an acceptable alternative. Please make sure that your instrument is capable of detecting this fluorochrome. Light sensitivity: This tandem dye is sensitive to photo-induced oxidation. Please protect this vial and stained samples from light. Fixation: Samples can be stored in IC Fixation Buffer (cat. 00-8222) (100 µL of cell sample + 100 µL of IC Fixation Buffer) or 1-step Fix/Lyse Solution (cat. 00-5333) for up to 3 days in the dark at 4°C with minimal impact on brightness and FRET efficiency/compensation. Some generalizations regarding fluorophore performance after fixation can be made, but clone specific performance should be determined empirically. Excitation: 488nm; Emission: 710 nm; Laser: Blue Laser. Filtration: 0.2 µm post-manufacturing filtered. The p19 shares sequence similarity with IL-6 subfamily members and is distantly related to the p35 subunit of IL-12. It shows no biological activity by itself; instead, it combines with the p40 subunit of IL-12 to form a biologically active, composite cytokine, IL-23. IL-23 shares some in vivo functions with IL-12, including the activation of the transcription factor Stat4. Similar to IL-12, human IL-23 stimulates IFN-g production and proliferation of PHA blast T cells, as well as in CD45 RO (memory) T cells. Ubiquitous transgenic expression of the IL-23 subunit p19 induces multiorgan inflammation, runting, infertility and premature death.
|4° C, store in dark, DO NOT FREEZE!|
|PBS with 0.1% gelatin and 0.09% sodium azide; pH 7.2|
For Research Use Only.
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