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Invitrogen™ 2'3'- Cyclic GAMP ELISA Kit

Product Code. 17174853
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Quantity:
96 Tests
Unit Size:
Pack of 96
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17174853 96 Tests Pack of 96
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Product Code. 17174853 Supplier Invitrogen™ Supplier No. EIAGAMP

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Description

ELISA

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.
TRUSTED_SUSTAINABILITY

Specifications

Accession Number 2'3 '-Cyclic GMP-AMP; cGAMP; Guanosine-Adenosine 2 ',3 '-cyclic monophosphate
Assay Range 0.04
Assay Sensitivity 0.08-20 pmol/mL
Conjugate HRP
Product Type ELISA
Sample Type Cell Lysates, Plasma, Supernatant, Tissue Homogenates
For Use With (Equipment) Colorimetric Microplate Reader
Interassay CV 8%
Intraassay CV 6
Kit Contents Anti-IgG coated 96 well plate, Standard, Assay Buffer Concentrate, Capture Antibody, HRP-conjugated Antigen, Wash Buffer Concentrate, TMB Substrate, Stop Solution, Plate Sealer
Label or Dye HRP
Quantity 96 Tests
Regulatory Status RUO
Sample Volume Cell Lysate 50 μL, Plasma 10 μL, Supernatant 50 μL, Tissue Homogenate 50 μL
Storage Requirements -20°C
Target cGAMP
Test Time 45 mins
Total Assay Time 2 hrs 30 mins
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