CCh, BaCl2, CTX, PGE2, ATP, acetazolamide and primaquine (Sigma-Aldrich, Steinheim, Germany) were dissolved in water. Forskolin and bumetanide (Sigma-Aldrich, Steinheim, Germany) were dissolved in ethyl alcohol, whilst Gly-H 101 (Merck, Darmstadt, Germany) and nocodazole (Sigma-Aldrich, Steinheim, Germany) were dissolved in DMSO. The Krebs buffer had the following composition in millimoles: NaCl 115.8, CaCl2 1.3, KCl 3.6, KH2PO4 1.4, NaHCO3 23.1 and MgSO4 1.2 (Merck, Darmstadt, Germany). The Krebs-mannitol buffer also contained Na-pyruvate (5.7 mM) (Sigma-Aldrich, Steinheim, Germany), Na-L-glutamate (5.1 mM) (Merck, Darmstadt, Germany) and D-mannitol (10 mM) (Sigma-Aldrich, Steinheim, Germany) and the Kreb-glucose buffer contained Na-pyruvate (5.7 mM), Na-L-glutamate (5.1 mM) and D-glucose (10 mM) (Sigma-Aldrich, Steinheim, Germany). In the chloride-free experiments, NaCl, KCl and CaCl2 were replaced with equimolar concentrations of Na-gluconate and K-gluconate and twice the concentration of Ca-gluconate (Sigma-Aldrich, Steinheim, Germany) to compensate for the chelation of calcium by gluconate. pH was set to 7.4 using acetic acid. In the bicarbonate-free experiments, NaHCO3 was replaced with equimolar concentration of NaH2PO4 (Merck, Darmstadt, Germany), the solution was gassed with 100 % O2 and the carbonic anhydrase was inhibited by acetazolamide (1 mM).
Acetazolamide
Manufactured by Merck Group
Sourced in United States, Germany, Italy, Macao
Acetazolamide is a chemical compound used as a laboratory reagent. It functions as a carbonic anhydrase inhibitor, which is a class of enzymes involved in various physiological processes. The core function of acetazolamide is to inhibit the activity of carbonic anhydrase enzymes, which can be utilized in various research and analytical applications.
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Lab products found in correlation
Amiloride, by Merck Group (4 mentions)
Bumetanide, by Merck Group (4 mentions)
Dimethyl sulfoxide (dmso), by Thermo Fisher Scientific (4 mentions)
Indomethacin, by Merck Group (3 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (3 mentions)
Lamotrigine, by Merck Group (2 mentions)
Topiramate, by Merck Group (2 mentions)
Milli q system, by Merck Group (2 mentions)
Theophylline, by Merck Group (2 mentions)
Benzenesulfonamide, by Merck Group (2 mentions)
Spr100, by Millar (2 mentions)
Pcu 2000 pressure control unit, by Millar (2 mentions)
Sulfanilamide, by Merck Group (2 mentions)
Methazolamide, by Merck Group (2 mentions)
Cisplatin, by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Primaquine, by Merck Group (2 mentions)
Nocodazole, by Merck Group (2 mentions)
47 protocols using acetazolamide
1
Ion Transport Modulation Assay
2
Modeling Fat Embolism Pathogenesis in Mice
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One hundred twenty C57BL/6J male mice weighing 22–24 g were purchased from Shanghai SLAC Laboratory Animal Co. Ltd. and used in the present study. Ninety-six mice were randomly divided into control and fat embolism groups 1–5 (n = 16 per group). Injectable fat was gathered from perinephric fat from allogeneic mice and homogenized to rupture the cell membrane, then centrifuged. Mice in the fat embolism groups were intravenously injected with 2 μL/g fat. Lung tissue was obtained 4, 6, 12, 24 and 48 h after fat injection. Sixteen mice were intravenously injected with the same volume of sterile saline as the control group. Pentobarbital at a dosage of 70 mg/kg was intraperitoneally injected as anesthesia. For protein function experiments, the AQP1 inhibitor bumetanide (Sigma, St. Louis, MO, USA) was dissolved in a NaOH liquor at a concentration of 2.25 g/L and intraperitoneally injected at a dose of 0.1 mg/g before fat injection, another AQP1 inhibitor Acetazolamide (Sigma) was intragastrically administered at a volume of 0.1 mL per mice (40 mg·kg−1·d−1), according to the manufacturer’s instructions.
3
Inhibition of Carbonic Anhydrase IX Activity
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Example 6
Anti-CA IX scFvs that are epitope mapped to the CA domain are able to block enzymatic activity. This is an important anti-tumor biological property of the antibodies that is distinct from their retargeting capacity. To evaluate the ability of anti-CA IX scFvs to block carbonic anhydrase activity, an assay is established to measure carbonic anhydrase activity of the CA IX paramagnetic proteoliposomes using established methods (Brion, 1988; Dodgson, 1991). In brief, the velocity of the reaction CO2+H2O ↔H2CO3 is measured by the time required for acidification of carbonate buffer in CO2 atmosphere, detected with phenol read as a pH indicator (Zavada, 1997). The reaction proceeds even in absence of the enzyme, with to=control time (this is set at 60 sec). Carbonic anhydrase reduces the time of acidification to t; one unit of the enzyme activity reduces the time to one half of control time: t/to=1. The CA IX paramagnetic proteoliposomes will be washed in PBS, resuspended in 1 mM carbonic buffer, pH 8.0. Acetazolamide (Sigma) serves as a positive control for inhibition of CA activity. Thus it is demonstrated that a purified anti-CA IX scFvs, in a dose dependent manner, blocks the CA activity of the proteoliposomes.
4
Pharmacological Compounds Acquisition Protocol
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Methanol, n-hexane, dichloromethane, ethyl acetate, n-butanol, hydrochlorothiazide (HCTZ), acetazolamide (ACTZ), N-ω-Nitro-L-arginine methyl ester (L-NAME), indomethacin, atropine, ouabain octahydrate, hexamethonium bromide, captopril, and propranolol were purchased locally from Sigma-Aldrich Chemicals. Acetonitrile and formic acid were obtained from J. T. Baker. All these chemicals were of standard analytical grade.
5
Intrathecal Administration of Neuroactive Compounds
6
Synthesis and Characterization of DTX-Loaded CMC Nanoparticles
7
Acetazolamide Sourcing and Usage
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Acetazolamide was purchased from Sigma Chemical Co. (St. Louis, MO, USA).
8
Preparing Pharmacological Compound Stocks
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Stocks of Rapamycin (R8781, Sigma, St. Louis, MO, USA), Retigabine (SML0325, Sigma), Minoxidil (M4145, Sigma), NS1643 (sc-2041353, Santa Cruz Biotechnology, Dallas, TX, USA), Lamotrigine (L3791, Sigma), Zolmitriptan (SML0248, Sigma), Cariporide (SML1360, Sigma), Topiramate (T0575, Sigma), Pantoprazole Sodium Hydrate (P0021, Sigma), Fenofibrate (F6020, Sigma), Acetazolamide (A6011, Sigma), Quercetin (Q4951, Sigma), Temozolomide (2706, Tocris, Bristol, UK), Dexamethasone (D4902, Sigma), ONO-RS-082 (O0766, Sigma), Topotecan hydrochloride (4562, Tocris), CKD 602 (5125, Tocris), (Z)-4-hydroxytamoxifen (3412, Tocris), Lansoprazole (2582, Tocris), Chlorzoxazone (C4397, Sigma), and Sodium Butyrate (B5887, Sigma), were made at 1000× concentration in DMSO. Stocks of Gabapentin (G154, Sigma) and Cisplatin (232120, EMD Millipore, Burlington, MA, USA) were made in water at 1000×. Dibutyryl cAMP sodium salt (D0627, Sigma) was the only compound dissolved directly in cell culture media at 1 mM concentration.
9
Detailed Synthesis of DTCs 1-31
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DTCs 1–30 were previosuly reported by one of our groups17 (link),18 (link) and were of > 99% purity. DTC 31 , acetazolamide, buffers and other reagents are commercially available from Sigma-Aldrich (Milan, Italy).
10
Astrocyte Culture and Immunodetection
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Unless otherwise noted, astrocytes were cultured in astrocyte media: DMEM (Life technologies, Austin TX) with 10% fetal calf serum (ATCC, VA). The following antibodies were utilized for studies: antiglial fibrillary acidic protein (Millipore AB5541, Darmstadt Germany, 1:500); antiaquaporin 4 (Millipore, AB3594, 1:200); anti-chicken alexa 568 (Thermofisher, A-11041, 1:200); anti-rabbit alexa 488 (Thermofisher, A-11008, 1:200). Acetazolamide was purchased from Sigma (St. Louis, MO).
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