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2 3 cgamp elisa

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The 2',3'-cGAMP ELISA is a laboratory assay used to detect and quantify the presence of the cyclic guanosine monophosphate-adenosine monophosphate (2',3'-cGAMP) molecule in biological samples. It is a sensitive and specific enzyme-linked immunosorbent assay (ELISA) that employs antibodies to capture and measure the target analyte.

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Lab products found in correlation

M per buffer, by Thermo Fisher Scientific (1 mentions) Pierce detergent compatible bradford assay, by Thermo Fisher Scientific (1 mentions) M per mammalian protein extraction reagent, by Thermo Fisher Scientific (1 mentions)

Close spelling variants of this product

2′3′-cGAMP ELISA Kit (36 citations) CGAMP ELISA (4 citations) 2′3′ cGAMP (3 citations) ELISAs (3 citations)

3 protocols using 2 3 cgamp elisa

1

Quantification of 2',3'-cGAMP in cells

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C129R and EP364R plasmids were transfected into PK-15 cells and PAMs and infected with ADV-GFP or HSV-GFP, respectively. In another experiment, 293-Dual hSTING-A162 cells were cotransfected with C129R and EP364R plasmids with 3×Flag-cGAS plasmid. PK-15 and 293-Dual hSTING-A162 cell lysates and cell supernatants were collected and subjected to ELISA for the determination of both the intracellular and extracellular 2′,3′-cGAMP level. A 2′,3′-cGAMP ELISA (Cayman Chemical; 501700) commercial kit was used for analysis according to the manufacturer’s instructions.
Dodantenna N., Ranathunga L., Chathuranga W.A., Weerawardhana A., Cha J.W., Subasinghe A., Gamage N., Haluwana D.K., Kim Y., Jheong W., Poo H, & Lee J.S. (2022). African Swine Fever Virus EP364R and C129R Target Cyclic GMP-AMP To Inhibit the cGAS-STING Signaling Pathway. Journal of Virology, 96(15), e01022-22.
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2

Quantifying Cellular cGAMP in Thymocytes

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To measure total cellular cGAMP DN3/4 thymocytes were expanded in OP9-DL1 culture for 3 days. Cells were then harvested, washed with PBS, and lysed in 100 µl MPER buffer (ThermoFisher Scientific) for 15 min at room temperature. Lysates were cleared by centrifugation at 16,000 × g for 15 min. Total protein content for lysates was determined using Pierce Detergent-Compatible Bradford Assay (ThermoFisher Scientific) for normalization of cGAMP. cGAMP ELISAs performed using the 2′,3′-cGAMP ELISA (Cayman Chemicals) according to the manufacturer’s recommended protocol.
Ratiu J.J., Barclay W.E., Lin E., Wang Q., Wellford S., Mehta N., Harnois M.J., DiPalma D., Roy S., Contreras A.V., Shinohara M.L., Wiest D, & Zhuang Y. (2022). Loss of Zfp335 triggers cGAS/STING-dependent apoptosis of post-β selection thymocytes. Nature Communications, 13, 5901.
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3

Quantification of 2'3'-cGAMP and IFN-λ2/λ3

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2’3’-cGAMP ELISA (Cayman) was performed according to manufacturer’s protocol. For cell quantification, cells were lysed in M-PER™ Mammalian Protein Extraction Reagent (Thermo Fisher). Cell lysate from each well was examined according to the manufacturer’s instructions.
For IFN-λ2/λ3 ELISA (R&D systems), plate was pre-coated with coating buffer overnight. After 3 washes with 1x Wash Buffer, plate was blocked with 1 x Reagent Dilute at room temperature for 1 hour. Standard samples or culture media from NSD1 WT or KO Cal27 cells were collected and incubated in the plate for 2 hours at room temperature. Plate was then washed 3 times with wash buffer. Detection antibody was diluted in Reagent Dilute and incubated in the plate for 2 hours at room temperature followed by 3 washes. Diluted Streptavidin-HRP B was added to each well and incubated at room temperature for 2 hours. Substrate solution was added and incubated in dark for 20 min. Plate was read at wavelength of 450 nm and 540 nm with a plate reader.
Li Y., Goldberg E.M., Chen X., Xu X., McGuire J.T., Leuzzi G., Karagiannis D., Tate T., Farhangdoost N., Horth C., Dai E., Li Z., Zhang Z., Izar B., Que J., Ciccia A., Majewski J., Yoon A.J., Ailles L., Mendelsohn C.L, & Lu C. (2022). Histone methylation antagonism drives tumor immune evasion in squamous cell carcinomas. Molecular cell, 82(20), 3901-3918.e7.
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