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Rabbit anti cgas

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Rabbit anti-cGAS is a primary antibody that binds to the cGAS (cyclic GMP-AMP synthase) protein. cGAS is a key sensor of cytosolic DNA and plays a critical role in the innate immune response. The antibody can be used to detect and study the expression and localization of cGAS in various cellular and experimental systems.

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Lab products found in correlation

Rabbit anti sting, by Cell Signaling Technology (5 mentions) Rabbit anti ptaok2, by GeneTex (2 mentions) Rabbit anti psting, by Cell Signaling Technology (2 mentions) Lsm 880, by Zeiss (2 mentions) Mouse anti gapdh, by Proteintech (2 mentions) Rabbit anti tbk1, by Cell Signaling Technology (2 mentions) Mouse anti neun, by Abcam (1 mentions) Goat anti iba1, by Abcam (1 mentions) Mouse anti gfap, by Cell Signaling Technology (1 mentions) Fv3000, by Olympus (1 mentions) Rabbit anti lc3 2, by Cell Signaling Technology (1 mentions) Rabbit anti ha, by Santa Cruz Biotechnology (1 mentions) Rabbit anti flag, by Santa Cruz Biotechnology (1 mentions) Mouse anti tubulin, by Merck Group (1 mentions) Mouse anti ha, by Cell Signaling Technology (1 mentions) Alexa fluor 488 anti rabbit igg, by Cell Signaling Technology (1 mentions) Mitotracker, by Thermo Fisher Scientific (1 mentions) Alexa fluor 488, by Abcam (1 mentions) Dapi fluoromount g, by Southern Biotech (1 mentions) Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)

Close spelling variants of this product

Anti-cGAS (29 citations) Rabbit monoclonal anti-cGAS (4 citations)

13 protocols using rabbit anti cgas

1

Immunohistochemical Analysis of Spinal Cord

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According to our previous report,63 (link) after being deeply anesthetized with pentobarbital sodium, rats were transcardially perfused with 150 mL of 1× PBS (4°C) followed by 150 mL of 4% paraformaldehyde (4°C). Lumbar enlargement of the spinal cord was collected, postfixed in 4% paraformaldehyde for 48 h at 4°C, and subsequently dehydrated with 30% sucrose for 3 days at 4°C. Subsequently, the spinal cord was cut transversely into slices with a thickness of 30 μm. The sections were permeabilized with 0.3% Triton X-100 for 10 min and blocked with 10% sheep or donkey serum for 2 h at 24°C. Sections were incubated with the following primary antibodies for 48 h at 4°C: goat-anti-Iba1 (1:200, Abcam, Cambridge, UK), mouse anti-GFAP (1:500, Cell Signaling Technology), mouse-anti-NeuN (1:500, Abcam), rabbit-anti-pTAOK2 (1:100, GeneTex), rabbit-anti-cGAS (1:100, Cell Signaling Technology), and rabbit-anti-STING (1:100, Cell Signaling Technology). After washing with 1× PBS, the sections were incubated with fluorescent secondary antibodies in the dark for 2 h at 24°C. Finally, the sections were examined under a fluorescence microscope (FV3000; Olympus, Japan).
Zhang H., Li A., Liu Y.F., Sun Z.M., Jin B.X., Lin J.P., Yang Y, & Yao Y.X. (2023). Spinal TAOK2 contributes to neuropathic pain via cGAS-STING activation in rats. iScience, 26(10), 107792.
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2

Western Blot Analysis of STING, cGAS, and Autophagy Markers

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Cell lysates were obtained after incubation in lysis buffer (1% Nonidet P-40, 150 mM NaCl, 50 mM Tris, pH 8.0) supplemented with complete protease inhibitor and resuspended in Laemmli buffer. Crude lysates were boiled for 5 m and then kept on ice. Proteins were separated by SDS-PAGE, transferred to PVDF membrane and incubated with 1 : 1000 rabbit anti-STING (Cell Signaling Technology, #13647S), 1 : 1000 rabbit anti-cGAS (Cell Signaling Technology, #15102S), 1 : 1000 rabbit anti-pSTING 1 : 1000 (Cell Signaling Technology, #85735S), rabbit anti-LC3-II (Cell Signaling Technology, #2775S) or 1 : 3000 mouse anti-LAMP1 (eBioscience, eBioH4A3) followed by 1 : 10 000 HRP-conjugated anti-rabbit or anti-mouse. Blots were developed using ECL™ Prime Western Blotting Detection Reagents.
Ng W.C., Kwek S.S., Sun B., Yousefi M., Ong E.Z., Tan H.C., Puschnik A.S., Chan K.R., Ooi Y.S, & Ooi E.E. (2022). A fast-growing dengue virus mutant reveals a dual role of STING in response to infection. Open Biology, 12(12), 220227.
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3

Western Blot Antibody Validation

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Commercial antibodies included rabbit anti-FLAG (Santa Cruz), rabbit anti-HA (Santa Cruz), mouse anti-tubulin (Sigma), mouse anti-HA (Cell Signaling Technology), rabbit anti-cGAS (Cell Signaling Technology), and mouse anti-STING (Proteintech). mouse anti-tubulin antibody was used for loading controls, and all mentioned antibodies were used at a dilution of 1:5,000 in 5% bovine serum albumin (BSA).
Qi S., Wang C., Zhang R., Zhu J., Hou X., Jiang Y., Li J., Ren M., Li M, & Hou F. (2022). Human STING Is Regulated by an Autoinhibitory Mechanism for Type I Interferon Production. Journal of Innate Immunity, 14(5), 518-531.
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4

Mitochondrial Dynamics in Activated CD4 T Cells

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1x106 CD4 T cells were stained with fluorochrome conjugated antibodies using a previously described method (Wang et al., 2021 (link)). For Top1/MitoTracker staining, HS CD4 T cells were treated with 10μM CPT (Cat# SKU: TG4110, TopoGEN, Buena Vista, CO) or DMSO (Cat# D2650, Sigma-Aldrich, St. Louis, MO) for 2 days. For cGAS/MitoTracker staining, HS-CD4 T cells were treated with 5μM CPT for 2 days. The primary antibodies included MitoTracker (Cat# M22425, Thermo Fisher Scientific, Waltham, MA), Rabbit anti-cGAS (Cat# 79978, Cell Signaling Technology, Danvers, MA), Rabbit anti-Top1mt (Cat# PA5-51660, Thermo Fisher Scientific, Waltham, MA) and mouse anti-Top1nc conjugated with Alexa Fluor 488 (Cat# ab223421, Abcam, Cambridge, MA). The secondary antibody included anti-rabbit IgG-Alexa Fluor 488 (Ca# 4412, Cell Signaling Technology). The cells were mounted with DAPI Fluoromount-G (Cat# D1306, SouthernBiotech, Birmingham, AL) and visualized with a confocal laser scanning inverted microscope (Leica Confocal, Model TCS sp8, Germany).
Dang X., Cao D., Zhao J., Schank M., Khanal S., Nguyen L.N., Wu X.Y., Zhang Y., Zhang J., Jiang Y., Ning S., Wang L., El Gazzar M., Moorman J.P, & Yao Z.Q. (2022). Mitochondrial topoisomerase 1 inhibition induces topological DNA damage and T cell dysfunction in patients with chronic viral infection. Frontiers in Cellular and Infection Microbiology, 12, 1026293.
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5

Visualizing HDAC3 and cGAS Colocalization

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To detect the colocalization of HDAC3 and cGAS, human primary macrophages were seeded on coverslips in 24-well plates. After transfection with HT-DNA, the cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.2% Triton X-100 for 10 min, and blocked in 3% BSA for 1 h. Cells were then incubated with primary antibodies (such as rabbit anti-cGAS (Cell Signaling Technology) and mouse anti-HDAC3 (Santa Cruz)) overnight at 4°C,. Alexa Fluor 488- and 546-conjugated secondary antibodies and Hoechst (Life technologies) were incubated for 1 h before the images were acquired using a ZEISS LSM 880 (Zeiss) confocal microscope.
To detect the localization of acetylated cGAS, PMA-differentiated FLAG-cGAS or FLAG-cGAS3KQ-expressing THP1 cells were seeded on coverslips in 24-well plates. After fixed with 4% paraformaldehyde for 15 min, permeabilized with 0.2% Triton X-100 for 10 min, and blocked in 3% BSA for 1 h, cells were then incubated with anti-FLAG antibodies overnight at 4°C. Alexa Fluor 546-conjugated secondary antibody and Hoechst (Life technologies) were incubated for 1 h before the images were acquired using a ZEISS LSM 880 (Zeiss) confocal microscope.
Dai J., Huang Y.J., He X., Zhao M., Wang X., Liu Z.S., Xue W., Cai H., Zhan X.Y., Huang S.Y., He K., Wang H., Wang N., Sang Z., Li T., Han Q.Y., Mao J., Diao X., Song N., Chen Y., Li W.H., Man J.H., Li A.L., Zhou T., Liu Z.G., Zhang X.M, & Li T. (2019). Acetylation Blocks cGAS Activity and Inhibits Self-DNA-Induced Autoimmunity. Cell, 176(6), 1447-1460.e14.
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6

Visualizing cGAS and G3BP1 in Stressed Cells

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Human primary macrophages, HeLa or U937 cells were seeded on coverslips in 24-well plates. After transfection with ISD or poly(I:C), or treatment with arsenite, the cells were fixed with 4% paraformaldehyde for 15 min, permeabilized with Triton X-100 for 10 min and blocked in 3% BSA for 1 h. Cells were then incubated with primary antibodies overnight at 4 °C, including rabbit anti-cGAS (Cell Signaling Technology) and mouse anti-G3BP1 (Abnova). Alexa Fluor 488- and 546-conjugated secondary antibodies (Life Technologies) were incubated for 1 h before images were acquired using a ZEISS LSM 880 (Zeiss) confocal microscope.
Liu Z.S., Cai H., Xue W., Wang M., Xia T., Li W.J., Xing J.Q., Zhao M., Huang Y.J., Chen S., Wu S.M., Wang X., Liu X., Pang X., Zhang Z.Y., Li T., Dai J., Dong F., Xia Q., Li A.L., Zhou T., Liu Z.G., Zhang X.M, & Li T. (2018). G3BP1 promotes DNA binding and activation of cGAS. Nature immunology, 20(1), 18-28.
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7

Immunofluorescence Staining of Fibroblasts

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Fibroblasts were grown on sterilized glass coverslips coated with 50 μg/ml fibronectin (F1141, Sigma‐Aldrich) and fixed with 4% paraformaldehyde in PBS for 20 min. Following fixation, cells were rinsed in PBS, permeabilized in PBS + 0.3% Triton X‐100 for 7 min, and then blocked in 10% FBS + PBS for 1 h. Both, primary and secondary antibodies were diluted in PBS + 0.05% Tween‐20 containing 5% FBS as follows. Primary antibodies: rabbit anti‐53BP1 (4937, Cell Signaling Technology), 1:100; mouse anti‐p21 (SC‐6246, Santa Cruz Biotechnology), 1:800; mouse anti‐Aurora B (Aim‐1; 611082, BD Biosciences), 1:500; rabbit anti‐cGAS (15102, Cell Signaling Technology), 1:200; mouse anti‐Hec1 (ab3613, Abcam), 1:1,500; mouse anti‐Plk1 (SC‐17783, Santa Cruz Biotechnology), 1:2,000; rabbit anti‐MCAK 49, 1:5,000; mouse anti‐α‐Tubulin (T5168, Sigma‐Aldrich), 1:1,500; human anti‐centromere antibody (ACA; kindly provided by Dr. W. C. Earnshaw), 1:3,000; rabbit anti‐Aurora B phospho T232 (600‐401‐677, ROCKLAND), 1:1,000; mouse anti‐Retinoblastoma (554136, BD Biosciences), 1:100. Secondary antibodies: Alexa Fluor‐488, Alexa Fluor‐568 and Alexa Fluor‐647 (Life Technologies), all 1:1,500. DNA was counterstained with 0.5 μg/ml DAPI (Sigma‐Aldrich) and coverslips mounted on slides with proper mounting solution.
Barroso‐Vilares M., Macedo J.C., Reis M., Warren J.D., Compton D, & Logarinho E. (2020). Small‐molecule inhibition of aging‐associated chromosomal instability delays cellular senescence. EMBO Reports, 21(5), e49248.
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8

Western Blot Analysis of TAOK2, cGAS, and STING

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The Western blot protocols were performed according to our previous studies.62 (link) After deep anesthesia with pentobarbital sodium, rats were decapitated, and lumbar enlargement of the spinal cord was quickly harvested and divided into four parts. The dorsal horns were selected and homogenized on ice using a lysis buffer containing protease and phosphatase inhibitors. The tissue homogenates were separated by 10% SDS-PAGE and subsequently transferred to 0.45-μm PVDF membranes. The membranes were blocked with 5% skimmed milk at 24°C for 1 h. Thereafter, membranes were incubated with the following primary antibodies for 24 h at 4°C: rabbit-anti-TAOK2 (1:500; GeneTex, TX, USA), rabbit-anti-pTAOK2 (1:1000, GeneTex), rabbit-anti-cGAS (1:1000, Cell Signaling Technology, Mass, USA), rabbit-anti-STING (1:500, Cell Signaling Technology), mouse-anti-tubulin (1:2000, Beyotime Biotechnology, Shanghai, China), and mouse-anti-GAPDH (1:10,000, Proteintech, IL, USA). After washing the primary antibody in TBST, the membrane was incubated with HRP-conjugated secondary antibody for 2 h at 24°C. The ChemiDoc MP System (Bio-Rad, Hercules, CA, USA) was used to detect the immune complex bands.
Zhang H., Li A., Liu Y.F., Sun Z.M., Jin B.X., Lin J.P., Yang Y, & Yao Y.X. (2023). Spinal TAOK2 contributes to neuropathic pain via cGAS-STING activation in rats. iScience, 26(10), 107792.
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9

Immunoblotting Analyses of DNA Damage Response

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Following antibodies were used: mouse anti beta-Actin (Abcam, ab6276, 1:1000), mouse anti-H2A.X phospho S139 (Millipore, 05-636, 1:1000), mouse anti-Histone H3 phospho-S10 (H3-pS10; Abcam, ab5176, 1:1000), rabbit anti-cleaved PARP (Cell Signaling, #9541, 1:1000), rabbit anti-PDL1 (Cell Signaling, #13684, 1:1000), rabbit anti-pTBK1 (Cell Signaling, #5483, 1:500), rabbit anti-TBK1 (Cell Signaling, #3504, 1:1000), rabbit anti-pSTING (Cell Signaling, #50907, 1:500), rabbit anti-STING (Cell Signaling, #13647, 1:1000), rabbit anti-cGAS (Cell Signaling, #15102, 1:1000), rabbit anti-MTH1 (Novus Biologicals, NB100-109, 1:1000). Secondary antibodies were: Peroxidase AffiniPure Donkey Anti-Rabbit IgG (Jackson ImmunoResearch, 711-035-152, 1:5000), Peroxidase AffiniPure Donkey Anti-Mouse IgG (Jackson ImmunoResearch, 715-035-150, 1:5000), IRDye 680RD Goat Anti-Mouse IgG (LI-COR, 926-68072, 1:5000) and IRDye 800CW Donkey Anti-Rabbit IgG (LI-COR, 926-32213, 1:5000).
Shen J., Guillén Mancina E., Chen S., Manolakou T., Gad H., Warpman Berglund U., Sanjiv K, & Helleday T. (2024). Mitotic MTH1 inhibitor TH1579 induces PD-L1 expression and inflammatory response through the cGAS-STING pathway. Oncogenesis, 13(1), 17.
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10

Western Blot Analysis of Sensorimotor Cortex

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7 days after surgery, the sensorimotor cortex adjacent to the infarcted core was dissected, and protein was extracted using RIPA buffer with PMSF (Beyotime, China). The protein levels were measured using the BCA protein assay kit (Vazyme, China) according to the manufacturer’s instructions. 30–50 μg protein per lane was loaded on SDS polyacrylamide gel and transferred to polyvinyl-difluoride membranes (Millipore, USA). The membranes were blocked using 5% non-fat milk or bovine serum albumin in Tris-buffered saline containing Tween 20 (TBST). Then, the membranes were incubated overnight at 4 ℃ with primary antibodies as follows: rabbit anti-STING (1:1000, Proteintech, 48,853), rabbit anti-TBK1 (1:1000, Cell Signaling Technology, 3504), rabbit anti-PTBK1 (1:1000, Abcam, ab109272), rabbit anti-cGAS (1:1000, Cell Signaling Technology, 31,659), mouse anti-GAPDH (1:10,000, Proteintech, 60,004–1-Ig), rabbit anti-Phospho-Stat1 (1:1000, Cell Signaling Technology, 9167), and rabbit anti-β-ACTIN (1:10,000, ABclonal, AC050). After incubation, the membranes were washed three times with TBST, and further incubated with HRP-conjugated goat anti-rabbit/mouse IgG secondary antibodies for 1 h at room temperature. After washing three times, the bands were detected using the enhanced chemiluminescence (ECL) advance kit (Vazyme, China) and quantified using ImageJ software (USA).
Wu C., Zhang S., Sun H., Li A., Hou F., Qi L, & Liao H. (2024). STING inhibition suppresses microglia-mediated synapses engulfment and alleviates motor functional deficits after stroke. Journal of Neuroinflammation, 21, 86.
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