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Trovafloxacin

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Trovafloxacin is a broad-spectrum antibiotic used in the laboratory setting. It is a member of the fluoroquinolone class of antibiotics and is effective against a wide range of bacteria, including both Gram-positive and Gram-negative organisms. Trovafloxacin functions by inhibiting the bacterial enzymes DNA gyrase and topoisomerase IV, which are essential for bacterial DNA replication and transcription.

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Lab products found in correlation

Spironolactone, by Merck Group (4 mentions) Probenecid, by Merck Group (3 mentions) Gsk269962, by Bio-Techne (2 mentions) Spermidine, by Merck Group (2 mentions) Dihydroxyacetone phosphate, by Merck Group (2 mentions) Inosine 5 monophosphate, by Merck Group (2 mentions) 7 aad, by Thermo Fisher Scientific (2 mentions) Udp glucose, by Abcam (2 mentions) Anti cd11b pe clone m1 70, by Thermo Fisher Scientific (2 mentions) Anti cd11c pe clone n418, by Thermo Fisher Scientific (2 mentions) Anti cd16 cd32 clone 93, by Thermo Fisher Scientific (2 mentions) Antibodies specific for mouse cd95, by BD (2 mentions) Human anti fas clone ch11, by Merck Group (2 mentions) Abt 737, by Abcam (2 mentions) Annexin 5 pacific blue, by BioLegend (2 mentions) Guanosine 5 monophosphate, by Merck Group (2 mentions) Fructose 1 6 bisphosphate, by Merck Group (2 mentions) Dexamethasone (dex), by Merck Group (2 mentions) Z vad fmk, by Enzo Life Sciences (2 mentions) Levofloxacin, by Merck Group (2 mentions)

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Trovafloxacin mesylate (3 citations)

13 protocols using trovafloxacin

1

P2X7 Receptor-Mediated Dye Uptake Assay

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Cells in poly-L-lysine-coated black optical 96-well plates (Nunc, 165305) were placed in PSS containing 0–2 mM Ca2+ and 1 µM YO-PRO1 (Invitrogen). The plate was transferred to a FLUOstar Omega reader (BMG Labtech) equilibrated to 37°C and 5% CO2. BzATP was injected to obtain a 0–500 µM final concentration (n=3) and fluorescence measured (4-mm orbital area) every 40 s for 30 min. Where indicated, cells were preincubated with 100 µM 10Panx (Tocris) for 10 min or 10–100 µg/ml trovafloxacin (Sigma) for 30 min and stimulated in the presence of inhibitors. After normalization for well-specific fluorescence, the average YO-PRO1 fluorescence of unstimulated cells was subtracted from that of agonist-stimulated cells to correct for bleaching. Dye uptake rates were derived by linear regression of data from the initial 5 min.
Adamczyk M., Griffiths R., Dewitt S., Knäuper V, & Aeschlimann D. (2015). P2X7 receptor activation regulates rapid unconventional export of transglutaminase-2. Journal of Cell Science, 128(24), 4615-4628.
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2

Multiparametric Flow Cytometry Analysis

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Trovafloxacin (Sigma-Aldrich, USA, PZ0015), GSK 269962 (Tocris bioscience, UK, 4009), Hoechst 33342 (Sigma-Aldrich, St Louis, MO). SYTO RNASelect green Fluorescent Cell Stain (S32703), MitoTracker Green (M7514), TO-PRO-3 (T3605) were purchased from Thermo Fisher Scientific. CD45-PeCy7 (557748; clone: HI30), CD3-APC (555335; clone: UCHT1), IgG1κ-PeCy7 (557872; clone: MOPC-21), A5-FITC (556419), A5-PE (556421), A5-APC (550474), A5-V450 (560506) and 10 × A5 binding buffer (556454) were purchased from BD Biosciences, CA. CD146-VioBlue (130-099-678; clone: 541-10B2), CD31-VioBlue (130-106-503; clone: AC128), CD45-FITC (130-080-202; clone: 5B1), CD14-FITC (130-080-701; clone: TÜK4), CD11b-FITC (130-081-201; clone: M1/70.15.11.5), IgG1-VioBlue (130-099-756), IgG2b-FITC (130-103-088), IgG2a-FITC (130-091-837), IgG1-FITC (130-098-847), IgG1-APC (130-098-846) were purchased from MAC Miltenyi Biotech, DE.
Jiang L., Paone S., Caruso S., Atkin-Smith G.K., Phan T.K., Hulett M.D, & Poon I.K. (2017). Determining the contents and cell origins of apoptotic bodies by flow cytometry. Scientific Reports, 7, 14444.
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3

Apoptosis Induction Assay Protocol

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Trovafloxacin, spironolactone, dexamethasone, spermidine, fructose 1,6-bisphosphate, dihydroxyacetone phosphate, inosine 5’-monophosphate, and guanosine 5’-monophosphate were obtained from Sigma. UDP-glucose was obtained from Abcam and Annexin V-Pacific Blue was from BioLegend. 7AAD, TO-PRO-3, anti-CD11b-PE (clone M1/70), anti-CD11c-PE (clone N418), and anti-CD16/CD32 (clone 93) were obtained from Invitrogen. Antibodies specific for mouse CD95 were obtained from BD. Human anti-Fas (clone CH11) was obtained from Millipore. Other reagents were obtained as follows: ABT-737 (abcam), TRAIL (Sigma), and zVAD-FMK (Enzo).
Medina C.B., Mehrotra P., Arandjelovic S., Perry J.S., Guo Y., Morioka S., Barron B., Walk S.F., Ghesquière B., Krupnick A.S., Lorenz U, & Ravichandran K.S. (2020). Metabolites released from apoptotic cells act as novel tissue messengers. Nature, 580(7801), 130-135.
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4

Apoptosis Induction Assay Protocol

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Reagents were obtained as follows: trovafloxacin, doxycycline and mitoxantrone (Sigma-Aldrich, MO), anti-Fas (clone CH11, Millipore, MA), annexin A5 (A5)-PE (BD Biosciences, CA) and TO-PRO-3 (Life Technologies, NY).
Tixeira R., Shi B., Parkes M.A., Hodge A.L., Caruso S., Hulett M.D., Baxter A.A., Phan T.K, & Poon I.K. (2018). Gasdermin E Does Not Limit Apoptotic Cell Disassembly by Promoting Early Onset of Secondary Necrosis in Jurkat T Cells and THP-1 Monocytes. Frontiers in Immunology, 9, 2842.
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5

Characterizing T Cell Responses to Viral Infection

Cited 1 time
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WT (CD4-Cre, ERT2-Cre-LSL-YFP), CD4-Cre Panx1fl/fl, ERT2-Cre-LSL-YFP-Panx1fl/fl, and CD4-Cre P2rx7fl/fl P14 cells were adoptively transferred into naive wild-type mice, which were infected with LCMV-Armstrong (2 × 105 PFU, intraperitoneally (i.p.)). Sometimes, wild-type (CD4-Cre, CMV-Cre), CMV-Cre Panx1fl/fl or CD4-Cre Panx1fl/fl mice were infected with LCMV Armstrong. In other experiments, wild-type (CD4-Cre) or CD4-Cre Panx1fl/fl mice were infected with Influenza-PR8 (1400 PFU, intranasally (i.n.)). In some experiments, LCMV-infected mice were treated with Trovafloxacin (4.2 mg x kg−1 mouse, Sigma-Aldrich) or Sodium Lactate (1.68 g x kg−1 mouse, Sigma-Aldrich)72 (link) between days 1–7 after infection.
Vardam-Kaur T., Banuelos A., Gabaldon-Parish M., Marie Wanhainen K., Hanqi Zhou M., van Dijk S., Gois Macedo B., Loureiro Salgado C., Santiago-Carvalho I., Peng C., Jameson S.C, & Borges da Silva H. (2023). Pannexin-1 channels promote CD8+ T cell effector and memory responses through distinct intracellular pathways. bioRxiv.
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6

Evaluating Hepatocyte Exposure to Drugs

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Troglitazone, nimesulide, caffeine, trovafloxacin, levofloxacin, LPS and methotrexate (all from Sigma, St Louis, Mo) were administered into the devices as 1% DMSO solutions in HMM at the final concentrations. To achieve testing of up to the recommended 100 × Cmax for in vitro liver systems, 1.0% DMSO final concentration was used to improve compound solubility. 1.0% DMSO is on the high end of published concentrations,7 (link),27 -29 (link) but the negative control, caffeine in 1.0% DMSO as well as DMSO alone, had no impact on the parameters measured in the devices. The flow was temporarily halted to allow imaging of each device using the GE IN Cell 6000 (GE LifeSciences, Piscataway NJ, USA) equipped with an environmental chamber to maintain 37° C and 5% CO2 (see below, HCA imaging and analysis). Flow was re-established in the devices after image collection.
Vernetti L.A., Senutovitch N., Boltz R., DeBiasio R., Ying Shun T., Gough A, & Taylor D.L. (2015). A human liver microphysiology platform for investigating physiology, drug safety, and disease models. Experimental Biology and Medicine, 241(1), 101-114.
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7

Apoptosis Induction Assay Protocol

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Trovafloxacin, spironolactone, dexamethasone, spermidine, fructose 1,6-bisphosphate, dihydroxyacetone phosphate, inosine 5’-monophosphate, and guanosine 5’-monophosphate were obtained from Sigma. UDP-glucose was obtained from Abcam and Annexin V-Pacific Blue was from BioLegend. 7AAD, TO-PRO-3, anti-CD11b-PE (clone M1/70), anti-CD11c-PE (clone N418), and anti-CD16/CD32 (clone 93) were obtained from Invitrogen. Antibodies specific for mouse CD95 were obtained from BD. Human anti-Fas (clone CH11) was obtained from Millipore. Other reagents were obtained as follows: ABT-737 (abcam), TRAIL (Sigma), and zVAD-FMK (Enzo).
Medina C.B., Mehrotra P., Arandjelovic S., Perry J.S., Guo Y., Morioka S., Barron B., Walk S.F., Ghesquière B., Krupnick A.S., Lorenz U, & Ravichandran K.S. (2020). Metabolites released from apoptotic cells act as novel tissue messengers. Nature, 580(7801), 130-135.
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8

Liposomal Transfection and Immune Stimulants

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DOTAP liposomal transfection reagent was from Roche (Mannheim, Germany). Cholera toxin B subunit (CTB) was from List Biological Laboratories, Inc. Ultra-pure lipopolysaccharide (E. coli O111:B4), ultra-pure lipopolysaccharide (S. minnesota RE595), poly(I:C) LMW and poly(dA:dT)/lyovec were from Invivogen. ATP, apyrase, carbenoxolone (CBX), bafliomycin A, brefeldin (BFA), 18-glycyrrhetinic (18GA), flufenamic acid (FFA), glibenclamide, gadolinium III (Gd3), probenecid, ARL67156 (an ecto-ATPase inhibitor) and trovafloxacin (a pannexin-1-selective antagonist) were from Sigma-Aldrich. Alum was from Thermo Scientific. Nigericin and Ac-DNLD-CHO (caspase-3/7 inhibitor) were from Calbiochem, and zVAD-FMK (pan-caspase-inhibitor) and zDEVD-FMK (caspase-3 inhibitor) was from R&D system. Fluorescent Yo-Pro-1 was from Life Technology.
Yang D., He Y., Muñoz-Planillo R., Liu Q, & Núñez G. (2015). Caspase-11 requires the pannexin-1 channel and the purinergic P2X7 pore to mediate pyroptosis and endotoxic shock. Immunity, 43(5), 923-932.
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9

Apoptosis Induction Assay Protocol

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Trovafloxacin, 7-aminoactinomycin D, cytochalasin D, sertraline, cycloheximide, serotonin and citalopram were obtained from Sigma-Aldrich (MO). Other reagents were obtained as follows: anti-Fas (05-201, 250 ng ml−1, clone CH11, Millipore, MA), annexin V–fluorescein isothiocyanate (FITC; BD Biosciences, CA), TO-PRO-3 (Life Technologies, NY), carboxyfluorescein diacetate succinimidyl ester (CFSE; Life Technologies) and GSK 269962 (Tocris bioscience, UK).
Atkin-Smith G.K., Tixeira R., Paone S., Mathivanan S., Collins C., Liem M., Goodall K.J., Ravichandran K.S., Hulett M.D, & Poon I.K. (2015). A novel mechanism of generating extracellular vesicles during apoptosis via a beads-on-a-string membrane structure. Nature Communications, 6, 7439.
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10

Fluorescence-based Antimicrobial Assay Protocol

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SYBR Green I (SGI) nucleic acid stain (10,000×; Lonza, Rockland, ME, USA) was stored at –20°C and thawed before use. A lysis buffer consisting of Tris (130 mM; pH 7.5), EDTA (10 mM), saponin (0.016%; W/V), and TritonX-100 (1.6%; V/V) was prepared in advance and stored at 4°C. Diminazene aceturate (DA; Novartis International AG, Basel, Switzerland), tetracycline hydrochloride, and chloroquine diphosphate (both drugs from Sigma-Aldrich Co., St Louis, MO, USA) were used as positive control drugs. Enrofloxacin (Figure 1A), enoxacin (Figure 1B), trovafloxacin (Figure 1C), norfloxacin (Figure 1D), ofloxacin (Figure 1E), luteolin, or pyronaridine tetraphosphate (PYR) (all drugs from Sigma-Aldrich Co.) was prepared as a 100 mM stock solution and stored at −20°C until use.
Rizk M.A., AbouLaila M., El-Sayed S.A., Guswanto A., Yokoyama N, & Igarashi I. (2018). Inhibitory effects of fluoroquinolone antibiotics on Babesia divergens and Babesia microti, blood parasites of veterinary and zoonotic importance. Infection and Drug Resistance, 11, 1605-1615.
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