Invitrogen™ 2'3'- Cyclic GAMP ELISA Kit

Description

The 2',3'-Cyclic GAMP Competitive ELISA Kit quantitates 2',3'-cyclic GAMP in lysed cells and tissue, tissue culture media, and EDTA plasma.∣Principle of the method: The 2',3'-Cyclic GAMP Competitive ELISA research-use-only kit is designed to quantitatively measure 2',3'-cyclic GAMP independent of species. A 2',3'-cyclic GAMP standard is provided to generate a standard curve for the assay and all samples are read off a user-generated standard curve. Standards or diluted samples are pipetted into a clear microtiter plate coated with anti-rabbit IgG antibodies. Anti-rabbit 2',3'-cyclic GAMP antibodies and 2',3'-cyclic GAMP-peroxidase conjugate is added to the wells. The 2',3'-cyclic GAMP-peroxidase conjugate and any 2',3'-cyclic GAMP in the sample will compete to bind to the anti-rabbit 2',3'-cyclic GAMP antibodies. After incubation, the plate is washed and substrate is added. The substrate reacts with the bound 2',3'-cyclic GAMP-peroxidase conjugate. After a 30 min incubation, the reaction is stopped and the intensity of the generated color is detected in a microtiter plate reader at 450 nm. The intensity of the generated color corresponds inversely to the amount of 2',3'-cyclic GAMP present in the sample.∣Rigorous validation: Each manufactured lot of this ELISA kit is quality tested for criteria such as sensitivity, specificity, precision, and lot-to-lot consistency. See manual for more information on validation.

Cyclic GMP-AMP (cGAMP) synthase (cGAS) is a nucleotidyltransferase located in the cytosol that acts as a cytosolic DNA sensor to detect foreign DNA from microbial pathogens as part of the innate immune response. Upon binding to cytosolic DNA, cGAS produces the cyclic dinucleotide second messenger cGAMP, which activates stimulator of interferon genes (STING), leading to activation of the type I interferon (IFN) pathway. In vitro, fibroblasts, macrophages, and dendritic cells isolated from cGAS knockout (cGAS-/-) mice do not produce type I IFNs following DNA transfection or DNA virus infection. Similarly, cells containing a frame-shift mutation in the cGAS locus fail to mount an immune response to HIV and other retroviruses. In vivo, cGAS-/- mice infected with herpes simplex virus 1 (HSV-1) have lower levels of IFN-α and IFN-β, shorter survival times, and higher post-mortem levels of HSV-1 in the brain.