2′,3′-Cyclic GAMP ELISA试剂盒背景:
2’,3’-Cyclic guanosine monophosphate–adenosine monophosphate (cyclic GMP-AMP, cGAMP, cyclic [G(2’,5’)pA(3’,5’)p]) was the first cyclic di-nucleotide found in metazoa1. 2’,3’-cGAMP is also referred to as “noncanonical” cGAMP due to the presence of the atypical 2’-5’ phosphodiester linkage between the guanosine and the adenosine. 2’,3’-Cyclic GAMP is a novel second messenger in innate immunity that regulates type I interferon (IFN) production1-6. Produced in mammalian cells by cGAS (cGAMP synthase) in response to double-stranded DNA in the cytoplasm binding to cGAS, cGAMP binds to the stimulator of interferon genes (STING). Subsequently STING induces the TBK1-IRF3-dependent production of IFN-β. This cGAS-cGAMP-STING pathway has been shown to play a critical role in pathogen detection and physiological conditions such as metabolic dysregulation, autoimmunity, and cancer7-10.Cellular concentrations of cGAMP are controlled by hydrolase enzymes that cleave the phospho-nucleotide bonds. One of these, ecto-nucleotide phosphatase, ENPP1, is a zinc-stimulated hydrolase of cGAMP and is present in cells, serum, and other samples11.
1. Wu, J., et al. (2012). Cyclic GMP-AMP is an endogenous second messenger in innate immune signaling by cytosolic DNA. Science, 339(6121), 826–30.
2. Sun, L., et al. (2013). Cyclic GMP-AMP synthase is a cytosolic DNA sensor that activates the type I interferon pathway. Science, 339(6121), 786–791.
3. Gao, P., et al. (2013). Cyclic [G(2’,5’)pA(3’,5’)p] is the metazoan second messenger produced by DNA-activated cyclic GMP-AMP synthase. Cell, 153(5), 1094–1107.
4. Ablasser, A., et al. (2013). cGAS produces a 2’-5’-linked cyclic dinucleotide second messenger that activates STING. Nature, 498(7454), 380–384.
5. Li, X-D., et al. (2013). Pivotal roles of cGAS-cGAMP signaling in antiviral defense and immune adjuvant effects. Science, 341(6152), 1390–1394.
6. Paijo, J. et al. (2016). cGAS senses human cytomegalovirus and induces type I interferon responses in human monocyte-derived cells. PLoS Pathogens, 12(4), e1005546.
7. Cai, X., et al. (2014). The cGAS-cGAMP-STING pathway of cytosolic DNA sensing and signaling. Molecular Cell 54(2), 289–296.
8. Guo, X., et al. (2017). Cyclic GMP-AMP ameliorales diet-induced metabolic dysregulation and regulates proinflammatory responses distinctly from STING activation. Nature, 7(6355), 1–13.
9. Gao, D., et al. (2015). Activation of cyclic GMP-AMP synthase by self-DNA causes autoimmune diseases.” Proceedings of the National Academy of Sciences, 112(42), E5699–E5705.
10. Bose, D. (2017). cGAS/STING pathway in cancer: Jekyll and Hyde story of cancer immune response. International Journal of Molecular Sciences, 18(11), 2456–2466.
11. Li, L., et al. (2014). Hydrolysis of 2’, 3’-cGAMP by ENPP1 and design of non-hydrolyzable analogs. Nature Chemical Biology, 10(12), 1043–1048.
艾美捷2′,3′-Cyclic GAMP ELISA试剂盒检测 #K067-H5/K067-H1原理:
The DetectX? 2’,3’-Cyclic GAMP (cGAMP) Immunoassay Kit is designed to quantitatively measure
2’,3’-cGAMP present in lysed cells and tissue, EDTA plasma, and tissue culture media samples.
Please read the complete kit insert before performing this assay. A 2’,3’-cGAMP standard is provided to generate a standard curve for the assay and all samples should be read off the standard curve. A clear microtiter plate coated with an antibody to capture rabbit IgG is provided. Standards or diluted samples are pipetted into the wells. A 2’,3’-cGAMP-peroxidase conjugate is added to the standards and samples in the wells. The binding reaction is initiated by the addition of a rabbit polyclonal antibody to 2’,3’-cGAMP to each well. After a 2 hour incubation, the plate is washed and substrate is added. The substrate reacts with the bound 2’,3’-cGAMP-peroxidase conjugate and after a short incubation the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader capable of measuring 450 nm wavelength. The concentration of the 2’,3’-cGAMP in the sample is calculated, after making suitable correction for the dilution of the sample, using software available with most plate readers.
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