Gsk 843

Manufactured by GlaxoSmithKline

The GSK 843 is a multipurpose laboratory equipment designed for various scientific applications. It functions as a versatile tool to assist researchers and scientists in their work, but a detailed description without interpretation or extrapolation is not available at this time.

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Lab products found in correlation

Lps eb ultrapure, by InvivoGen (2 mentions) Gsk 872, by GlaxoSmithKline (2 mentions) Gsk 963, by GlaxoSmithKline (2 mentions) Itaconate, by Merck Group (1 mentions) Dimethyl malonate, by Merck Group (1 mentions) 4 octyl itaconate, by Merck Group (1 mentions) Necrostatin 1, by Merck Group (1 mentions) Cl264, by InvivoGen (1 mentions) Interleukin 4 (il 4), by Thermo Fisher Scientific (1 mentions) Gm csf, by BioLegend (1 mentions) Z yvad fmk, by Enzo Life Sciences (1 mentions) Necrostatin 1 nec 1, by Enzo Life Sciences (1 mentions) Mntbap, by Enzo Life Sciences (1 mentions) Purified lps, by InvivoGen (1 mentions) Trolox, by Enzo Life Sciences (1 mentions) Caspase 3 9662, by Cell Signaling Technology (1 mentions) Mlkl mabc604, by Merck Group (1 mentions) Ripk1 38 rip, by BD (1 mentions) Anti flag m2, by Merck Group (1 mentions) Anti fkbp12, by Thermo Fisher Scientific (1 mentions)

4 protocols using gsk 843

Investigating Cell Signaling Pathways

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The following chemical reagents were used in cell culture experiments: LPS-EB Ultrapure (1 μg/ml, Invivogen), zVAD (50 μm, SM Biochemicals), GSK 963 (100nM, GlaxoSmithKline), GSK 843 (100nM, GlaxoSmithKline), GSK 872 (100nM, GlaxoSmithKline), AP1 (100 nM, ClonTech, sold as “B/B Homodimerizer”), itaconate (1mM, Sigma), citrate (1mM, Sigma), dimethyl malonate (1mM, Sigma), 4-octyl itaconate (1mM, provided by Luke O’Neill, Trinity College, Dublin). For in vivo intracranial injections, the following concentrations were used: GSK 963 and GSK 843, 800nM; dimethyl malonate and 4-octyl itaconate, 8mM.

Daniels B.P., Kofman S.B., Smith J.R., Norris G.T., Snyder A.G., Kolb J.P., Gao X., Locasale J.W., Martinez J., Gale M J.r., Loo Y.M, & Oberst A. (2019). The nucleotide sensor ZBP1 and kinase RIPK3 induce the enzyme IRG1 to promote an antiviral metabolic state in neurons. Immunity, 50(1), 64-76.e4.

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Cell Culture Experiments with PRR Ligands

Cited 1 time

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The following PRR ligands and chemical inhibitors were used in cell culture experiments: Poly(I:C) (1 μg/ml, Millipore), LPS-EB Ultrapure (1 μg/ml, Invivogen), CL264 (1 μg/ml, Invivogen), zVAD (50 μM, SM Biochemicals), necrostatin-1 (30 μM, Sigma), GSK 963 (100nM, GlaxoSmithKline), GSK 843 (100nM, GlaxoSmithKline) (Mandal et al., 2014 (link)), GSK 872 (100nM, GlaxoSmithKline) and QVD (20 μM, SM Biochemicals), AP1 (100 nM, ClonTech, sold as “BB Homodimerizer”).

Daniels B.P., Snyder A.G., Olsen T.M., Orozco S., Oguin TH I.I.I., Tait S.W., Martinez J., Gale M J.r., Loo Y.M, & Oberst A. (2017). RIPK3 restricts viral pathogenesis via cell death-independent neuroinflammation. Cell, 169(2), 301-313.e11.

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Generating Bone Marrow-Derived Cells

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For BMDCs, 10 × 10⁶ BM cells were plated in 10 cm tissue culture dish and cultured for a week with 10 ng/ml GM-CSF (BioLegend) and 5 ng/ml IL-4 (Biosource) in RPMI1640 media supplemented with 10% FCS, 2 mM Glutamine, 100 units/ml penicillin, and 100 μg/ml streptomycin. Fresh GM-CSF and IL-4 were added on day 4. Floating cells were collected, plated out and subjected to subsequent experimental analyses. For BMDMs, 10 × 10⁶ BM cells were plated in 10 cm tissue culture dish and cultured for a week in L929-conditioned media. Fresh L929-conditioned media were added on day 4. Attached macrophages were plated out and subjected to subsequent experimental analyses. Purified LPS from Invivogen was used for all experiments. In some experiments, BMDCs were pretreated with RIPK3 kinase inhibitor GSK’843 (GlaxoSmithKline) (Kaiser et al., 2013 (link)), Necrostatin-1 (Nec-1) (Enzo life sciences), z-YVAD-fmk (Enzo life sciences), N-acetyl cysteine (NAC) (Calbiochem), MnTBAP (Enzo life sciences), and Trolox (Enzo life sciences) for an hour prior to LPS stimulation. After stimulation, culture media and cells were used for ELISA, RNA, and protein analyses.

Moriwaki K., Balaji S., McQuade T., Malhotra N., Kang J, & Chan F.K. (2014). The Necroptosis Adaptor RIPK3 Promotes Injury-Induced Cytokine Expression and Tissue Repair. Immunity, 41(4), 567-578.

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Multimodal Analysis of Cell Death Pathways

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Where indicated, the following drugs were used at the listed concentrations: 50 μM zVAD (SM Biochemicals), 100nM GSK’843 (GlaxoSmithKline).
The following antibodies were used for Western Blots and immunoprecipitations: ADAR1 (15.8.6) SantaCruz, ZBP1 (Zippy-1) AdipoGen, actin (13E5) Cell Signaling Technology, MDA5 (D74E4) Cell Signaling Technologies, p-RIPK3 (GEN135–35-9), Genentech, RIPK3 (1G6.1.4) Genentech, or RIPK3 (2283) ProSci, p-MLKL (D6E3G) Cell Signaling Technologies, MLKL (MABC604) Millipore, Caspase-3 (9662) Cell Signaling Technologies, Cleaved Capsase-3 (9661) Cell Signaling Technologies, RIPK1 (38/RIP) BD Biosciences, anti-FLAG (M2) Sigma, anti-FKBP12, Thermo Fisher (PA1–026A). Iba1 (Cat no. 019–19741) Wako-Chem, Cleaved Caspase 3 (Clone D3E9) Cell Signaling Technologies were used in immunohistochemical analysis.
The following antibodies were used for flow cytometry analysis of splenocytes: FITC anti-CD19 (clone 1D3; BD Biosciences) PerCP-Cy5.5 anti-CD3e (clone 145–2C11; BD Biosciences), PE-Cy7 anti-Ly6C (clone HK1.4; Biolegend), APC anti-F4/80 (clone BM8; eBioscience), AF700 anti-Ly6G (clone 1A8; Biolegend), APC-Cy7 anti-NK1.1 (clone PK136; BD Biosciences), BV510 anti-CD8a (clone 53–6.7; BD Biosciences); BV605 anti-CD4 (clone RM4–5; BD Biosciences), BV650 anti-CD11b (clone M1/70; Biolegend), and BUV395 anti-CD45.2 (clone 104; BD Biosciences).

Hubbard N.W., Ames J.M., Maurano M., Chu L.H., Somfleth K.Y., Gokhale N.S., Werner M., Snyder J.M., Lichauco K., Savan R., Stetson D.B, & Oberst A. (2022). ADAR1 mutation causes ZBP1-dependent immunopathology. Nature, 607(7920), 769-775.

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