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CartChicken IL-12/IL-23 p40 Do-It-Yourself ELISA, ≤10 Plates
Chicken IL-12 p40 ELISA Data
Chicken IL-12 p40 ELISA Kit Components
| Component | Usage | Quantity | Catalog # |
| Anti-Chicken IL-12/IL-23 p40 Polyclonal Antibody | Capture Antibody | 100 µg | PB0435C-100 |
| Biotinylated Anti-Chicken IL-12/IL-23 p40 Polyclonal Antibody | Detection Antibody | 50 µg | PBB0436C-050 |
| Chicken IL-12/IL-23 p40 Recombinant Protein Standard | Standard | 5 µg | RP0289C-005 |
Chicken IL-12 p40 ELISA Specifications
The Chicken IL-12/IL-23 p40 Do-It-Yourself ELISA contains capture antibody, standard, and detection antibody for development of a Chicken IL-12/IL-23 p40 ELISA. The antibodies have been determined to function in an ELISA with the standard provided. Optimal buffers, concentrations, incubation times, incubation temperatures, and methods for the ELISA have not been determined. A working knowledge of ELISA is strongly recommended. The quantities of components provided are not matched. Components may also be purchased separately.
For additional tips and techniques to ensure a successful ELISA, check out our ELISA Technical Guide.
IL-12 Background
IL-12 is a member of the IL-12 family, which includes IL-12, IL-23, IL-27, and IL-35. Like other IL-12 family members, IL-12 is a heterodimeric cytokine. IL-12 is encoded by two separate genes, IL-12A (p35) and IL-12B (p40). The active heterodimer, and a homodimer of p40 are formed following protein synthesis. IL-12 is involved in the differentiation of naive T cells into Th1 cells. It is known as a T cell-stimulating factor, which can stimulate the growth and function of T cells. It stimulates the production of IFN-γ and TNF-α from T and natural killer (NK) cells, and reduces IL-4 mediated suppression of IFN-γ.
Alternate Names - IL12B, CLMF, CLMF2, IL-12B, IMD28, NKSF, NKSF2, IMD29, Interleukin 12 subunit beta, interleukin 12B
Chlamydia psittaci PmpD-N Exacerbated Chicken Macrophage Function by Triggering Th2 Polarization and the TLR2/MyD88/NF-κB Signaling Pathway
Chu J, Li X, Qu G, Wang Y, Li Q, Guo Y, Hou L, Liu J, Eko FO, He C.
Int J Mol Sci. 2020 Mar 15;21(6):2003. doi: 10.3390/ijms21062003.
Applications: Measurement of chicken IL-2, IL-6, IL-10, IL-12, and IFN-γ in cell culture supernatants by ELISA
Abstract
The polymorphic membrane protein D (PmpD) is a highly conserved outer membrane protein which plays an important role in pathogenesis during Chlamydia psittaci infection. In this study, we evaluated the ability of the N-terminus of PmpD (PmpD-N) to modulate the functions of chicken macrophages and the signaling pathway(s) involved in PmpD-N-induced Toll-like receptors (TLRs), as well as interleukin (IL)-6 and IL-10 cytokine secretions. Thus, HD11 macrophages were treated with exogenous and intracellular PmpD-N of C. psittaci. The chlamydial growth was evaluated by enumeration of chlamydial loads in the infected macrophages. The phagocytic function of macrophages following PmpD-N treatment was detected by fluorescein-labeled Escherichia coli (E. coli). The concentration of nitric oxide (NO) secreted by HD11 macrophages was measured by the amount of NO2- in the culture supernatant using the Griess method. The cytokine secretions were assessed using multiplex cytokine ELISA kits. Expression levels of TLRs, myeloid differentiation factor 88 (MyD88), and nuclear factor kappa B (NF-κB) were analyzed by a Western blotting assay, as well as a luciferase assay, while NF-κB p65 nuclear translocation was assessed by confocal microscopy. The nuclear translocation of the transcription factor NF-κB was confirmed by evaluating its ability to combine with the corresponding promoter using the electrophoretic mobility shift assay (EMSA). After treatment with exogenous or endogenous PmpD-N, chlamydial loads and phagocytic functions were reduced significantly compared with those of the plasmid vector group, while NO secretions were reduced significantly compared with those of the lipopolysaccharide (LPS) treatment. Stimulation of HD11 cells with PmpD-N provoked the secretion of the Th2 cytokines, IL-6, and IL-10 and upregulated the expression of TLR2, TLR4, MyD88, and NF-κB. Furthermore, inhibition of TLR2, MyD88, and NF-κB in HD11 cells significantly decreased IL-6 and IL-10 cytokine levels, while NO production and phagocytosis increased significantly, strongly suggesting their involvement in PmpD-N-induced Th2 cytokine secretion and macrophage dysfunction. Our data indicate that C. psittaci PmpD-N inhibited macrophage functions by activating the Th2 immune response and the TLR2/MyD88/NF-κB signaling pathway.
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