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Meclofenamate

Manufactured by Merck Group
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Meclofenamate is a non-steroidal anti-inflammatory drug (NSAID) used as a laboratory reagent. It functions as an inhibitor of cyclooxygenase (COX) enzymes, which are involved in the production of prostaglandins. Meclofenamate can be used in various in vitro and in vivo research applications to study the effects of COX inhibition on biological processes.

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Lab products found in correlation

Microvue complement c3a plus, by Quidel (3 mentions) Edta vacutainer tube, by BD (2 mentions) Lomustine, by Merck Group (1 mentions) Txb2 eia kit, by Cayman Chemical (1 mentions) Tonabersat, by Merck Group (1 mentions) Latrunculin a, by Merck Group (1 mentions) Calcein am, by Merck Group (1 mentions) Brefeldin a, by Merck Group (1 mentions) Thromboxane b2 eia kit, by Cayman Chemical (1 mentions) Anti inhibitory kappa bα ikbα sc 371, by Santa Cruz Biotechnology (1 mentions) Ctad vacutainer tubes, by BD (1 mentions) Asserachrom β tg, by Diagnostica Stago (1 mentions) Txb2 elisa kit, by Cayman Chemical (1 mentions) Benzamidine, by Merck Group (1 mentions) Arachidonic acid, by Merck Group (1 mentions) Miconazole, by Merck Group (1 mentions) Iberiotoxin, by Merck Group (1 mentions) Capsaicin, by Merck Group (1 mentions) Emodepside, by Bayer (1 mentions) Ctad tubes, by BD (1 mentions)

10 protocols using meclofenamate

1

Preparation of Meclofenamate and Lomustine

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Meclofenamate and lomustine were purchased from Sigma. For preparation of 100-mM stock solutions, Meclofenamate was dissolved in dimethyl sulfoxide (DMSO) and lomustine in ethanol. Both drugs were aliquoted and stored at −20 °C. Final DMSO and ethanol concentrations did not exceed 0.1%.
Schneider M., Potthoff A.L., Evert B.O., Dicks M., Ehrentraut D., Dolf A., Schmidt E.N., Schäfer N., Borger V., Pietsch T., Westhoff M.A., Güresir E., Waha A., Vatter H., Heiland D.H., Schuss P, & Herrlinger U. (2021). Inhibition of Intercellular Cytosolic Traffic via Gap Junctions Reinforces Lomustine-Induced Toxicity in Glioblastoma Independent of MGMT Promoter Methylation Status. Pharmaceuticals, 14(3), 195.
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2

Measuring Coagulation and Inflammation Markers

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Beta-thromboglobulin (βTG) and prothrombin fragments 1+2 (F1+2) were measured by commercial enzyme-linked immunosorbent assay (ELISA) in plasma samples collected in CTAD tubes as previously described (24 (link)). EDTA plasma was used to measure histamine (ELISA kit Starfish, Gernusco, Italy) and C3a levels. Because the manufacturer of C3a kits recently modified their ELISA to the C3a Plus version (Microvue Complement C3a Plus, Quidel, San Diego, CA), archived GalTKO.hCD46 samples were reanalyzed for this study. Blood, collected at 3 time points (0, 15 and 60min) in EDTA tubes containing 100μl (at 10μg/ml) of meclofenamate (Sigma-Aldrich, St.Louis, MO), was analyzed to measure plasma levels of thromboxane B2 (TXB2 EIA Kit, Cayman Chemical Company, Ann Arbor, MI; Catalog No. 519031).
Burdorf L., Riner A., Rybak E., Salles I., De Meyer S., Shah A., Quinn K., Harris D., Zhang T., Parsell D., Ali F., Schwartz E., Kang E., Cheng X., Sievert E., Zhao Y., Braileanu G., Phelps C., Ayares D., Deckmyn H., Pierson R I.I.I, & Azimzadeh A. (2016). Platelet sequestration and activation during GalTKO.hCD46 pig lung perfusion by human blood is primarily mediated by GPIb, GPIIb/IIIa, and VWF. Xenotransplantation, 23(3), 222-236.
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3

Compound Preparation for Cell Assays

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Meclofenamate (10 mM in water), tonabersat (40 mM in DMSO), latrunculin A (1 mM in DMSO), brefeldin A (10 mM in DMSO), and calcein AM (4 mM in DMSO), were purchased from Sigma Aldrich.
Gava F., Rigal L., Mondesert O., Pesce E., Ducommun B, & Lobjois V. (2018). Gap junctions contribute to anchorage-independent clustering of breast cancer cells. BMC Cancer, 18, 221.
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4

Measuring Inflammatory and Coagulation Markers

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Beta-thromboglobulin (βTG) and prothrombin fragments 1+2 (F1+2) were measured by commercial enzyme-linked immunosorbent assay (ELISA) in plasma samples collected in CTAD tubes as previously described (14 (link)). EDTA plasma was used to measure histamine (ELISA kit Starfish, Gernusco, Italy) and C3a levels (14 (link)). Blood, collected at 3 time points (0′, 15′ and 60′) in EDTA tubes containing 100 μl (at 10 μg/ml) of meclofenamate (Sigma-Aldrich, St. Louis, MO), was analyzed to measure plasma levels of thromboxane (Thromboxane B2 EIA Kit, Cayman Chemical Company, Ann Arbor, MI; Catalog No. 519031).
Burdorf L., Stoddard T., Zhang T., Rybak E., Riner A., Avon C., Laaris A., Cheng X., Sievert E., Braileanu G., Newton A., Phelps C.J., Ayares D., Azimzadeh A.M, & Pierson RN I.I.I. (2014). Expression of human CD46 modulates inflammation associated with GalTKO lung xenograft injury. American journal of transplantation : official journal of the American Society of Transplantation and the American Society of Transplant Surgeons, 14(5), 1084-1095.
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5

NF-κB Signaling Pathway Antibodies

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Phospho-specific anti-IκBα (serine 32/36, #9246) and anti-p65 (serine 536, #3036S) antibodies were purchased from Cell Signaling Technology Inc. (Danvers, MA, USA). Anti-β-actin (sc-8432), anti-NF-κB p65 (sc-8008), and anti-inhibitory kappa Bα (IkBα) (sc-371) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-nuclear matrix protein p84 (ab-487) was purchased from Abcam (Cambridge, MA, USA). Goat Anti-mouse IgG (#401215) or Goat Anti-rabbit IgG (#401315) was purchased from Calbiochem (Carlsbad, CA, USA) and used as the secondary antibody. Other chemicals used in the current experiment, including meclofenamate, were purchased from Sigma-Aldrich (St. Louis, MO, USA).
Ryu J., Kim K.I., Hossain R., Lee M., Hong J.T., Lee H.J, & Lee C.J. (2023). Meclofenamate Suppresses MUC5AC Mucin Gene Expression by Regulating the NF-kB Signaling Pathway in Human Pulmonary Mucoepidermoid NCI-H292 Cells. Biomolecules & Therapeutics, 31(3), 306-311.
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6

Plasma Biomarkers for Thrombosis

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For βTG, blood perfusate was collected in CTAD vacutainer tubes (Becton Dickinson, Franklin Lakes, NJ, USA) after breaking the vacuum and left on ice for 20 min until centrifugation at 1430g for 20 minutes. For TXB2, blood perfusate was collected in EDTA vacutainer tubes (Becton Dickinson) supplemented with meclofenamate (10μg/mL) (Sigma-Aldrich) and immediately centrifuged at 514g for 15 minutes. Plasma was collected and stored at −80C until assayed. Assays for beta-thromboglobulin (βTG; Asserachrom β-TG, Diagnostica Stago, Asnieres, France) and thromboxane B2 (TXB2; TXB2 ELISA Kit, Cayman Chemicals, Ann Arbor, MI, USA) were performed according to manufacturer’s instructions. The normal reference ranges for human βTG and TXB2 are <50 IU/mL and 1–2 pg/mL respectively.
Cimeno A., Hassanein W., French B.M., Powell J.M., Burdorf L., Goloubeva O., Cheng X., Parsell D.M., Ramsoondar J., Kuravi K., Vaught T., Uluer M.C., Redding E., O’Neill N., Laird C., Hershfeld A., Tatarov I., Thomas K., Ayares D., Azimzadeh A.M., Pierson RN I.I.I., Barth R.N, & LaMattina J.C. (2017). N-glycolylneuraminic acid knock out reduces erythrocyte sequestration and thromboxane elaboration in an ex vivo pig-to-human xenoperfusion model. Xenotransplantation, 24(6), 10.1111/xen.12339.
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7

Blood Biomarker Sampling During Liver Perfusion

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Blood was collected from the suprahepatic vena cava outflow at timed intervals throughout the perfusion in EDTA vacutainer tubes (Becton Dickinson), supplemented with meclofenamate (10μg/mL) and benzamidine (1mM) (Sigma-Aldrich, St. Louis, MO, USA) after the vacuum was broken. The tubes were centrifuged at 2500rpm for 15 minutes. Plasma was stored at −80°C until used. Alanine aminotransferase (ALT) and aspartate aminotransferase (AST) levels were measured via commercial laboratory (Antech, Rockville, MD, USA).
Cimeno A., French B.M., Powell J.M., Phelps C., Ayares D., O’Neill N.A., Laird C.T., Pierson RN I.I.I., Azimzadeh A.M., Barth R.N, & LaMattina J.C. (2017). Synthetic Liver Function is Detectable in Transgenic Porcine Livers Perfused with Human Blood. Xenotransplantation, 25(1), 10.1111/xen.12361.
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8

Plasma Biomarkers of Thrombosis and Inflammation

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Beta-thromboglobulin (βTG), prothrombin fragments 1+2 (F1+2), were measured by commercial enzyme-linked immunosorbent assay (ELISA) in plasma samples collected in CTAD tubes as previously described (31 EDTA plasma was used to measure histamine (ELISA kit Starfish) and C3a levels (Microvue Complement C3a Plus, Quidel) (29 (link)). Blood, collected in EDTA tubes containing 100μl of meclofenamate (10μg/ml, Sigma-Aldrich), was used for thromboxane (Thromboxane B2 EIA Kit).
Burdorf L., Harris D., Dahi S., Zhang T., Ali F., Shah A., Thompson M., Braileanu G., Cheng X., Sievert E., Schwartz E., Sendil S., Parsell D.M., Redding E., Phelps C.J., Ayares D.L., Azimzadeh A.M, & Pierson RN 3.r.d. (2018). THROMBOXANE AND HISTAMINE MEDIATE PVR ELEVATION DURING XENOGENEIC PIG LUNG PERFUSION WITH HUMAN BLOOD. Xenotransplantation, 26(2), e12458.
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9

Emodepside and Neurophysiological Agents

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Emodepside used in this study was obtained from Bayer Animal Health. Diethylcarbamazine (DEC), 4-aminopyridine, miconazole, meclofenamate, arachidonic acid, iberiotoxin, and capsaicin were obtained from Sigma Aldrich (St. Louis, MO, USA). The drugs were dissolved in distilled water or DMSO and diluted in recording solution to obtain final concentration.
Verma S., Kashyap S.S., Robertson A.P, & Martin R.J. (2020). Diethylcarbamazine activates TRP channels including TRP-2 in filaria, Brugia malayi. Communications Biology, 3, 398.
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10

Biomarkers of Thrombosis and Inflammation

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Beta-thromboglobulin (βTG) and prothrombin fragments 1+2 (F1+2), were measured by commercial enzymelinked immunosorbent assay (ELISA) using plasma samples collected in CTAD tubes (Becton Dickinson, Franklin Lakes, NK, USA). EDTA plasma was used to measure histamine (ELISA kit Starfish) and C3a levels (Microvue Complement C3a Plus, Quidel). Blood, collected in EDTA tubes containing 100 μL of meclofenamate (10 μg/ml, Sigma-Aldrich), was used for thromboxane (Thromboxane B2 EIA Kit).
Additionally, human blood serum samples were tested for anti-Neu5Gc IgG antibodies using Maxisorp 96-well flat bottom plates (Nunc, cat #44-2404-21) coated with 1 μg/ml of Neu5Gc-PAA (Glycotech, cat #08-051) or PAA (Glycotech, cat #08-000). Affinity purified horseradish peroxidase (HRP) -conjugated goat anti-human IgG (Jackson, cat #109-035-008) was used to detect the anti-Neu5Gc IgG antibodies. All ELISA plates were measured using Spectramax M3 (Molecular Devices).
Chaban R., Habibabady Z., Hassanein W., Connolly M.R., Burdorf L., Redding E., Laird C., Ranek J., Braileanu G., Sendil S., Cheng X., Sun W., O'Neill N.A., Kuravi K., Hurh S., Ayares D.L., Azimzadeh A.M, & Pierson RN 3.r.d. (2022). Knock-out of N-glycolylneuraminic acid attenuates antibody-mediated rejection in xenogenically perfused porcine lungs. Xenotransplantation, 29(6).
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