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- Assay Type Competitive ELISA
- Sample Types Cell Lysates, Tissue Extracts, Tissue Culture Media
- Sensitivity 0.048 pmol/mL
- Species 2',3'-Cyclic GAMP is identical across species
- Assay Duration 2.5 Hours
- Samples/Plate 39 in Duplicate
- Readout Colorimetric, 450 nm
- Standard Curve
The 2′,3′-Cyclic GAMP ELISA Kit quantitatively measures 2’,3’-cGAMP present in lysed cells and tissue, EDTA plasma, and tissue culture media samples. The 2′,3′-Cyclic GAMP ELISA Kit is a competitive ELISA with a run time of 2.5 hours. Please read the complete kit insert before performing this assay.
Use our provided 2’,3’-Cyclic GAMP (cGAMP) standard to generate a standard curve for the assay. Pipette the standards or diluted samples into a transparent microtiter plate coated with our goat anti-rabbit IgG antibody. Add the 2’,3’-cGAMP-peroxidase conjugate and the 2’,3’-cGAMP polyclonal rabbit antibody. Then incubate the mixture for 2 hours shaking at room temperature. The immunological reaction occurs between the anti-2’,3’-cGAMP polyclonal antibody, the 2’,3’-cGAMP antigen in the sample or standard, and the 2’,3’-cGAMP-peroxidase conjugate. As the 2’,3’-cGAMP concentration in the sample increases, the bound 2’,3’-cGAMP-peroxidase conjugate decreases, causing a decrease in signal and vice versa.
After the 2-hour incubation, wash away the excess 2’,3’-cGAMP-peroxidase and add the TMB substrate. The TMB substrate reacts with the bound 2’,3’-cGAMP-peroxidase conjugate generating a signal detected by a plate reader at 450nm. Use the intensity and the standard curve to calculate the 2’,3’-cGAMP concentration in the samples.
2’,3’-Cyclic guanosine monophosphate–adenosine monophosphate (2’,3’-cGAMP)is a mammalian messenger molecule. 2’,3’-cGAMP was the first cyclic di-nucleotide found in metazoa. 2’,3’-cGAMP is a “noncanonical“ cGAMP due to 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) production. 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 is critical in pathogen detection and physiological conditions such as metabolic dysregulation, autoimmunity, and cancer.