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2 3 cgamp elisa kit

Manufactured by Cayman Chemical
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The 2′3′-cGAMP ELISA Kit is a quantitative assay designed to measure the concentration of 2′3′-cGAMP, a cyclic dinucleotide, in biological samples. The kit utilizes a competitive enzyme-linked immunosorbent assay (ELISA) format.

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Lab products found in correlation

M per mammalian protein extraction reagent, by Thermo Fisher Scientific (5 mentions) Lysis buffer, by Thermo Fisher Scientific (2 mentions) M pertm mammalian protein extraction reagent, by Thermo Fisher Scientific (2 mentions) Lipofectamine 2000, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions) Ripa buffer, by Thermo Fisher Scientific (2 mentions) M per mammalian protein extraction reagent lysis buffer, by Thermo Fisher Scientific (1 mentions) Lipofectamine 2000 reagent, by Thermo Fisher Scientific (1 mentions) Click it plus edu alexa fluor 647 flow cytometry assay kit, by Thermo Fisher Scientific (1 mentions) Synergy h1 hybrid microplate reader, by Agilent Technologies (1 mentions) Celltiter glo luminescent cell viability assay, by Promega (1 mentions) Ufc8010, by Merck Group (1 mentions) M per buffer, by Thermo Fisher Scientific (1 mentions) Human ifn beta duoset elisa, by R&D Systems (1 mentions) Duoset elisa, by R&D Systems (1 mentions) Mouse ifn alpha all subtype elisa kit, by PBL Assay Science (1 mentions) Lumikine xpress mifn β 2, by InvivoGen (1 mentions) M per lysis buffer, by Thermo Fisher Scientific (1 mentions) Ripa lysis and extraction buffer, by Thermo Fisher Scientific (1 mentions) Duoset elisa development system, by R&D Systems (1 mentions)

Close spelling variants of this product

2′,3′-cGAMP ELISA 2′3′ cGAMP CGAMP ELISA kit CGAMP ELISA ELISAs

36 protocols using 2 3 cgamp elisa kit

1

Cytokine and Cyclic Nucleotide Quantification

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Cells supernatant were collected, and the expression of IFN-β protein was detected by human, and porcine IFN-β ELISA kit (Solarbio), according to the manufacturer’s instructions. Cells infected with FMDV, SVV, or EV-A71 were collected, and the level of 2’3’-cGAMP was detected by 2’3’-cGAMP ELISA kit (Cayman Chemical Co., Ann Arbor, MI), according to the manufacturer’s instructions.
Liu H., Zhu Z., Xue Q., Yang F., Li Z., Xue Z., Cao W., He J., Guo J., Liu X., Shaw A.E., King D.P, & Zheng H. (2023). Innate sensing of picornavirus infection involves cGAS-STING-mediated antiviral responses triggered by mitochondrial DNA release. PLOS Pathogens, 19(2), e1011132.
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2

Quantifying cGAMP Levels in Cells

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For cGAMP measurements, 1 × 106 TRAMP-C2 and A549 cells were grown in a culture dish and treated with 4 μg/ml ISD or water as control for 18 h. Cells were collected and lysed using M-PER mammalian protein extraction reagent. After lysed cells were spun down, supernatant was collected and cGAMP levels were measured using the 2′3′-cGAMP ELISA kit according to manufacturer’s instructions (Cayman Chemical, USA).
Suter M.A., Tan N.Y., Thiam C.H., Khatoo M., MacAry P.A., Angeli V., Gasser S, & Zhang Y.L. (2021). cGAS–STING cytosolic DNA sensing pathway is suppressed by JAK2-STAT3 in tumor cells. Scientific Reports, 11, 7243.
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3

Quantifying cGAMP Production

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cGAMP production was measured using the 2’3’-cGAMP ELISA kit from Cayman Chemicals (cat # 501700-96) according to the manufacturer’s instructions.
MacDonald K.M., Nicholson-Puthenveedu S., Tageldein M.M., Khasnis S., Arrowsmith C.H, & Harding S.M. (2023). Antecedent chromatin organization determines cGAS recruitment to ruptured micronuclei. Nature Communications, 14, 556.
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4

Quantification of cGAMP in Transfected Cells

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4 × 106 or 6 × 106 U2OS cells (for TREX1 KO and PML KO experiments, respectively) were seeded into 15-cm dishes, and 24 h later cells were transfected with the Bluescript vector (empty vector control) and Human L1 plasmid (used in L1 retrotransposition assay) using Lipofectamine 2000 reagent (Invitrogen). Cells were harvested 36 h after transfection, washed with 2× with PBS and pelleted before lysis. Samples were resuspended in 500 μl M-PER (mammalian protein extraction reagent) lysis buffer (Thermo Scientific). Lysates were incubated on ice for 30 min with gentle agitation every 10 min, before being spun down at 16 000 × g, 4°C for 10 min. Samples were quantified using the 2′3′cGAMP ELISA Kit (Cayman Chemical) according to the manufacturer's instructions.
Mathavarajah S., Vergunst K.L., Habib E.B., Williams S.K., He R., Maliougina M., Park M., Salsman J., Roy S., Braasch I., Roger A.J., Langelaan D.N, & Dellaire G. (2023). PML and PML-like exonucleases restrict retrotransposons in jawed vertebrates. Nucleic Acids Research, 51(7), 3185-3204.
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5

Multimodal Analysis of Cellular Responses

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EdU incorporation assay was performed with Click-iT™ Plus EdU Alexa Fluor™ 647 Flow Cytometry Assay Kit (Thermo Fisher Scientific, Cat# C10634), following the manufacturer’s instructions. Cells were plated in triplicates in 6-well plates and incubated with 10 μM EdU for 1.5–2 h, followed by flow cytometric analysis.
For cell viability assays, H2087-LCC cells were plated in four replicates in 96-well plates and incubated with 100 pM TGF-β1 or 2 μM CDK4/6 inhibitor palbociclib (Selleckchem, Cat# S1116) in culture media. Cell proliferation was quantified at the indicated time points using the CellTiter-Glo® Luminescent Cell Viability Assay (Promega, Cat# G7572), following the manufacturer’s instructions. Samples were analyzed using the Synergy H1 Hybrid microplate reader (Agilent).
For determinations of 2’3’-cGAMP, 5 × 105 cells were lysed using M-PER™ mammalian protein extraction reagent (Thermo Fisher Scientific, Cat# 78503). 2’3’-cGAMP content was measured with a 2’3’-cGAMP ELISA kit (Cayman Chemical Company, Cat# 501700) following the manufacturer’s instructions.
Hu J., Sánchez-Rivera F.J., Wang Z., Johnson G.N., Ho Y.J., Ganesh K., Umeda S., Gan S., Mujal A., Delconte R., Hampton J.P., Zhao H., Kottapalli S., de Stanchina E., Iacobuzio-Donahue C.A., Pe’er D., Lowe S.W., Sun J.C, & Massagué J. (2023). STING inhibits the reactivation of dormant metastasis in lung adenocarcinoma. Nature, 616(7958), 806-813.
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6

Quantifying cGAMP in THP1 Cells

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In all, 2 × 106 cells THP1 cells were treated with 1 μM dox for 1 h and subsequently harvested by centrifugation to measure cGAMP concentrations using a 2′3′-cGAMP ELISA Kit (Cayman Cat# 501700) in accordance with the manufacturer’s protocol.
Banerjee D., Langberg K., Abbas S., Odermatt E., Yerramothu P., Volaric M., Reidenbach M.A., Krentz K.J., Rubinstein C.D., Brautigan D.L., Abbas T., Gelfand B.D., Ambati J, & Kerur N. (2021). A non-canonical, interferon-independent signaling activity of cGAMP triggers DNA damage response signaling. Nature Communications, 12, 6207.
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7

Quantifying 2′3′-cGAMP Hydrolysis Activity

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To determine 2′3′-cGAMP hydrolysis activity of recombinant proteins, 1 μL of each recombinant protein (2 μM) and 1 μL of 2′3′-cGAMP (21 μM) were added to 8 μL of buffer (50 mM HEPES-KOH pH 7.5, 40 mM KCl, 1 mM DTT). Reactions were carried out at 37 °C for 1 h. Then, each mixture was diluted to 3 mL and transferred to a 10 kDa ultrafiltration tube (Millipore, UFC8010), followed by centrifugation at 4000 g for 5 min at 4 °C. After centrifugation, 50 μL of each supernatant was used to assay the amount of 2′3′-cGAMP with the 2′3′-cGAMP ELISA Kit (Cayman, 501700), according to the manufacturer’s instructions. Each test was performed three times. Results were analyzed statistically via two-tailed Student’s t test.
Yin M., Kuang W., Wang Q., Wang X., Yuan C., Lin Z., Zhang H., Deng F., Jiang H., Gong P., Zou Z., Hu Z, & Wang M. (2022). Dual roles and evolutionary implications of P26/poxin in antagonizing intracellular cGAS-STING and extracellular melanization immunity. Nature Communications, 13, 6934.
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8

Quantifying 2'3'-cGAMP in Transfected Cells

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HeLa LT and U-2-OS were seeded in 12-well plates, transfected with siRNA and 24 h later cells were split and seeded in 6-well plates. For stimulation with dsDNA90, cells were seeded in 6-well plates and transfected with 4 μg/ml using Lipofectamine 2000 (Thermo Fisher Scientific). 72 h after siRNA treatment and 24 h after dsDNA90 transfection, cells were harvested, counted, washed with PBS, pelleted, and stored at −80°C. To quantify 2′3′-cGAMP levels, pellets corresponding to 250,000 cells (U-2-OS) or 500,000 cells (HeLa LT) were resuspended in 130 μL M-PER buffer (Thermo Fisher Scientific), incubated on ice for 15 min, centrifuged at 16,000 g at 4°C and 2′3′-cGAMP levels were quantified using the 2′3′-cGAMP ELISA Kit (Cayman Chemical) in technical duplicate according to the manufacturer’s instructions.
Segura-Bayona S., Villamor-Payà M., Attolini C.S., Koenig L.M., Sanchiz-Calvo M., Boulton S.J, & Stracker T.H. (2020). Tousled-Like Kinases Suppress Innate Immune Signaling Triggered by Alternative Lengthening of Telomeres. Cell Reports, 32(5), 107983.
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9

Quantifying 2'3'-cGAMP in Cells

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3 × 105 cells were seeded onto a 6-well plate and lyzed after 36-hour culture. The cellular 2’3’-cGAMP levels were measured using a 2’3’-cGAMP ELISA kit (Cayman) in accordance with the manufacturer’s instructions.
Wang Y., Luo M., Chen Y., Wang Y., Zhang B., Ren Z., Bao L., Wang Y., Wang J.E., Fu Y.X., Luo W, & Wang Y. (2020). ZMYND8 expression in breast cancer cells blocks T-lymphocyte surveillance to promote tumor growth. Cancer research, 81(1), 174-186.
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10

Measuring cGAMP Levels in Cell Lysates

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cGAMP concentration in cell lysates was measured using a 2’3’-cGAMP ELISA Kit (Cayman Chemical, U.S.A.) according to the manufacturer’s protocol. The result was normalized by total protein concentration as described previously [16 (link)].
Ueda K., Sakai C., Ishida T., Morita K., Kobayashi Y., Horikoshi Y., Baba A., Okazaki Y., Yoshizumi M., Tashiro S, & Ishida M. (2023). Cigarette smoke induces mitochondrial DNA damage and activates cGAS-STING pathway: application to a biomarker for atherosclerosis. Clinical Science (London, England : 1979), 137(2), 163-180.
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