zLEHD-fmk, zIETD-fmk and zVAD-fmk were obtained from Enzo Life Sciences (Lörrach, Germany). Rotenone, 3-nitropropionic acid (3-NP), antimycin A, KCN, oligomycin, FB1 and H2O2 were purchased from Sigma-Aldrich.
3 nitropropionic acid 3 np
Manufactured by Merck Group
Sourced in United States
3-nitropropionic acid (3-NP) is a chemical compound that is used as a laboratory reagent. It is a white crystalline solid that is soluble in water and organic solvents. 3-NP is commonly used in research applications, but its specific function and intended use should not be extrapolated or interpreted.
Automatically generated - may contain errors
Lab products found in correlation
Rotenone, by Merck Group (2 mentions)
Drp1 sirna, by Bioneer (2 mentions)
Mitotracker probe, by Thermo Fisher Scientific (2 mentions)
Oligomycin, by Merck Group (1 mentions)
Zlehd fmk, by Enzo Life Sciences (1 mentions)
Z ietd fmk, by Enzo Life Sciences (1 mentions)
Antimycin a, by Merck Group (1 mentions)
Z vad fmk, by Enzo Life Sciences (1 mentions)
Ana 12, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide mtt, by Merck Group (1 mentions)
Brain derived neurotrophic factor (bdnf), by Alomone (1 mentions)
Digitonin, by Merck Group (1 mentions)
Clark type oxygen electrode, by Oroboros (1 mentions)
N acetylcysteine, by Merck Group (1 mentions)
Scrambled sirna, by Horizon Discovery (1 mentions)
Hoechst 33342 dye, by Thermo Fisher Scientific (1 mentions)
2 3 5 triphenyltetrazolium chloride ttc, by Merck Group (1 mentions)
Nrf2 small interfering rna sirna, by Santa Cruz Biotechnology (1 mentions)
Mouse monoclonal antibody against β actin, by Proteintech (1 mentions)
Ml385, by MedChemExpress (1 mentions)
11 protocols using 3 nitropropionic acid 3 np
1
Apoptosis Induction Inhibitors Protocol
2
Neuroinflammation and Neurodegeneration Assays
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TrkB inhibitor (ANA-12, N-[2-[[(Hexahydro-2-oxo-1H-azepin-3-yl)amino]carbonyl]phenyl]-benzo[b]thiophene-2-carboxamide Cat#SML0209) and 3-nitropropionic acid (3NP, Cat#N22908) 3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT, Cat#M5655) were obtained from Sigma-Aldrich Argentina. Ultrapure bacterial lipopolysaccharide (LPS) (Escherichia coli, 0111:B4, Cat#tlrl-3pelps) was from Invivogen (USA); BDNF was purchased from Alomone (Cat#B-250). Culture media and supplements were acquired from Invitrogen Argentina. All other media and supplements were obtained from Sigma-Aldrich Argentina, unless otherwise specified.
3
Mitochondrial Respiration Profiling
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Complex-dependent mitochondrial respiration was measured using a Clark-type oxygen electrode (OROBOROS, Innsbruck, Austria) as previously described 32 . Briefly, approximately 0.5 million cells were permeabilized with 0.02 mg/ml digitonin (Sigma-Aldrich) and added into the chamber with substrate-free respiration buffer. After recording the baseline, CI-, CII-, and CI+CII-dependent respiration were measured by adding malate (1.6 mM) + glutamate (8 mM), succinate (4 mM), or malate (1.6 mM) + glutamate (8 mM) + succinate (4 mM), respectively. Rotenone (0.8 μM, Sigma-Aldrich), 3-nitropropionic acid (3-NP, 12.6 mM, Sigma-Aldrich), and Rotenone (0.8 μM) + 3-NP (12.6 mM, Sigma-Aldrich) were used as inhibitors for CI-, CII-, and CI+CII- dependent respiration, respectively. All respiration data were normalized with cell numbers.
4
Mitochondrial Dynamics Regulation Protocols
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The YFP-fused mito-tracker plasmid (pmito-YFP) was previously described [28] (link). 3-nitropropionic acid (3-NP) and N-acetylcysteine (NAC) were purchased from Sigma (St. Louis, MO USA). Mdivi-1 (3-(2,4-Dichloro-5-methoxyphenyl)-2,3-dihydro-2-thioxo-4(1H)-quinazolinone) was purchased from Enzo life sciences (Farmingdale, NY, USA). A mitoTracker probe was purchased from Invitrogen (Carlsbad, CA). The validated siRNA targeting for Cpn10 (#1, 5′-CAAAGUAGUUCUAGAUGAC-3′), (#2, 5′-GCGUGAAAGUUGGAGAUAA-3′) negative scrambled siRNA (5′-CCUACGCCACCAAUUUCGU-3′) were purchased form Dharmacon (Thermo Scientific). And previously validated Drp1 siRNA (5′-GAGGUUAUUGAACGACUCA-3′) and Opa1 siRNA (5′-CUGGAAAGACUAGUGUGUU-3′) were synthesized from Bioneer (Daejeon, Korea).
5
Plasmid Constructs and Reagents for Cellular Imaging
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YFP-fused mito-tracker plasmid (pmito-YFP) was previously described [32] . GFP-fused hnRNP A1 plasmid (pEGFP-hnRNP A1) was purchased from Origene Technologies (Rockville, MD). 3-nitropropionic acid (3-NP) was purchased from Sigma (St. Louis, MO). A mitoTracker® probe and Hoechst 33,342 dye were purchased from Invitrogen (Carlsbad, CA). The validated siRNAs targeting for hnRNP A1 (#1, 5′-GCUCUUCAUUGGAGGGUUG-3′), (#2, 5′-CUACAAUGAUUUUGGGAAUUU-3′) and a negative scrambled siRNA (5′-CCUACGCCACCAAUUUCGU-3′) were purchased from Dharmacon (Thermo Scientific). Previously validated Drp1 siRNA (5′-GAGGUUAUUGAACGACUCA-3′) was synthesized from Bioneer (Daejeon, Korea) [32] .
6
Rat model of 3NP-induced brain injury
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All experimental procedures were conducted according to the guidelines of the Institutional Animal Care and Use Committee at Konkuk University, Seoul, Korea. Female Sprague-Dawley rats (n=10) weighing 300–320 g were used in this study. 3-nitropropionic acid (3NP; Sigma-Aldrich, St. Louis, MO, USA) was dissolved in saline solution, and the pH was adjusted to 7.4 with NaOH. Animals received a subcutaneous injection of 3NP (15 mg/kg) for 4 weeks, once every two days. Animals were anesthetized with chloral hydrate (400 mg/kg, intraperitoneally) and sacrificed 2 days after the final injection. Control rats received the equivalent volume of normal saline solution. To evaluate tissue injury by 3NP, brains (n=3) were quickly removed and were sliced at 1-mm thickness. Brain slices were incubated at 37°C for 30 min in a 2% solution of 2,3,5-triphenyltetrazolium chloride (TTC; Sigma-Aldrich).
7
Suspending DMF and Dissolving 3-NP
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DMF was suspended in 0.08% carboxymethyl cellulose (CMC), and 3-nitropropionic acid (3-NP) was dissolved in saline; both were purchased from Sigma-Aldrich (MO, United States). All other chemicals were of analytical grade, and the used kits and antibodies sources are provided in the relevant methodology section.
8
Neuroprotective Evaluation of β-BA
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The 3-nitropropionic acid (3-NP) was purchased from Sigma-Aldrich (St. Louis, MO, USA). β-BA was provided as an ex gratia sample from BAPEX, Rajasthan. Analytical-grade versions of all other compounds were also employed in the investigation. Drug and chemical solutions were freshly prepared before administration to the rats. The recommended dosage of 3-NP is 10 mg/kg intraperitoneally (i.p.), given for 15 days. The saline solution contained 5% DMSO and had a pH of 7.4. For oral use, β-BA was dissolved in water (with 2% ethanol) (p.o.). As a standard drug, vitamin E (Evion® 200 marketed formulation, manufactured by Merck Limited, New York City, NY, USA, manufacturing date: May 2018 and expiry date: July 2020) was dissolved in water (with ethanol) and administered by the oral route (p.o.).
9
Tolfenamic Acid and Huntingtin Regulation
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Tolfenamic acid (purity ≥98%) was purchased from a commercial supplier (Abcam, Cambridge, MA, USA), and 3-nitropropionic acid (3-NP) was purchased from Sigma-Aldrich (MO, USA). The mouse monoclonal antibody against huntingtin (EM48) was purchased from Millipore (CA, USA). The rabbit polyclonal antibodies against LC3, P62, and HO1; the mouse monoclonal antibody against β-actin; and horseradish peroxidase-conjugated secondary antibodies were purchased from Proteintech (Wuhan, China). ML385, a specific Nrf2 inhibitor, was purchased from MedChem Express (NJ, USA). Nrf2 small interfering RNA (siRNA) and rabbit polyclonal antibody against Sp1 were purchased from Santa Cruz Biotechnology Inc. (CA, USA). The rabbit polyclonal antibody against NQO1 and D,L-buthionine-(S,R)-sulfoximine (BSO), a specific glutathione synthase inhibitor, was purchased from Abcam (Cambridge, MA, USA).
10
Huntington's Disease Induction Protocol
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The experimental model to induce HD was previously described [10 (link)21 (link)]. Briefly, 3-nitropropionic acid (3NP; Sigma, St. Louis, MO, USA) was dissolved in saline (25 mg/mL) and passed through a 0.2-µm filter. We followed the 3NP injection protocol described by Huang et al. [21 (link)], with 3NP injected intraperitoneally twice daily for two days at 12-h intervals (10:00 a.m. and 10:00 p.m.) at a dose of 60 mg/kg for the first two injections and 80 mg/kg for the second two injections.
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