Specifications

  • Catalog Number K067-H1D
  • 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 183 in Duplicate
  • Readout Colorimetric, 450 nm

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Assay Principle:

The DetectX® 2′,3′-Cyclic GAMP ELISA Kit (384-Well Plate) quantitatively measures 2’,3’-cGAMP present in lysed cells and tissue, EDTA plasma, and tissue culture media samples. This competitive ELISA has 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.

Protocol Summary:

  • Pipette standards or diluted samples into the provided transparent microtiter plate, pre-coated with goat anti-rabbit IgG antibody.
  • Add 2’,3’-cGAMP-peroxidase conjugate and 2’,3’-cGAMP polyclonal rabbit antibody to initiate the immunological reaction.
  • Incubate the mixture shaking at room temperature for 2 hours. The reaction varies inversely with the 2’,3’-cGAMP concentration in the sample.
  • After incubation, remove excess conjugate and introduce the TMB substrate. This substrate reacts with the bound conjugate, producing a measurable signal.
  • Use a plate reader to detect the signal at 450nm and calculate the 2’,3’-cGAMP concentration based on the standard curve.

Background:

2’,3’-Cyclic guanosine monophosphate–adenosine monophosphate (2’,3’-cGAMP) is a significant mammalian messenger molecule, first discovered as a cyclic di-nucleotide in metazoa. It is known as a “noncanonical“ cGAMP due to its unusual 2’-5’ phosphodiester linkage between guanosine and adenosine. As a novel second messenger in innate immunity, 2’,3’-cGAMP plays an important role in regulating type I interferon (IFN) production. In mammalian cells, cGAMP is synthesized by cGAS (cGAMP synthase) in response to cytoplasmic double-stranded DNA binding to cGAS. Once produced, cGAMP activates the stimulator of interferon genes (STING), leading to the TBK1-IRF3-dependent production of IFN-β.

The cGAS-cGAMP-STING pathway is not only crucial in pathogen detection but also influences metabolic dysregulation, autoimmunity, and cancer. Understanding the dynamics of this pathway and its molecular components may lead to new therapeutic strategies in immunology and oncology.

The DetectX® 2′,3′-Cyclic GAMP ELISA Kit (384-Well Plate) offers sensitive and specific quantification of 2′,3′-cGAMP. This kit is valuable for researchers studying cellular responses to DNA stimuli and the broader implications of 2′,3′-cGAMP signaling in health and disease.

2′,3′-Cyclic GAMP ELISA Kit (384-Well Plate)

$1,698.00

In Stock

The DetectX® 2′,3′-cGAMP ELISA Kit measures 2′,3′-cGAMP in tissues and cells.

SKU: K067-H1D Category:

Research Areas