6 hydroxydopamine 6 ohda

Manufactured by Merck Group

Sourced in United States, Japan

6-hydroxydopamine (6-OHDA) is a chemical compound used in research laboratories. It is a selective neurotoxin that is primarily used to induce dopaminergic neurodegeneration in experimental models. The core function of 6-OHDA is to selectively target and damage dopamine-producing neurons, which can be useful for studying Parkinson's disease and other neurological disorders.

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Lab products found in correlation

6 ohda, by Merck Group (6 mentions) Desipramine, by Merck Group (4 mentions) Apomorphine, by Merck Group (4 mentions) Ascorbic acid, by Merck Group (4 mentions) L ascorbic acid, by Merck Group (3 mentions) β actin, by Merck Group (2 mentions) Lipopolysaccharides (lps), by Merck Group (2 mentions) Stereotaxic frame, by Kopf Instruments (2 mentions) Stereotaxic frame, by Narishige (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Sh sy5y cell, by American Type Culture Collection (2 mentions) Suramin, by Merck Group (1 mentions) Cox 2, by Cayman Chemical (1 mentions) Caspase 3, by Novus Biologicals (1 mentions) Nf κb, by Merck Group (1 mentions) Phospho extracellular signal regulated kinase p erk, by Cell Signaling Technology (1 mentions) Anti akt, by Cell Signaling Technology (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Anti p akt, by Cell Signaling Technology (1 mentions) Anti pten, by Cell Signaling Technology (1 mentions)

Spelling variants of this product

6-OHDA (353 citations) 6-hydroxydopamine (28 citations) 6-hydroxydopamine hydrobromide (6-OHDA, (11 citations) 6-hydroxydopamine hydrochloride (6-OHDA) (4 citations) 6-hydroxydopamine (6-OHDA; H4381, (3 citations)

80 protocols using 6 hydroxydopamine 6 ohda

Neuroinflammation Markers Profiling Protocol

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6-OHDA (6-hydroxydopamine; Sigma, St. Louis, MO, USA; catalog: H4381)

β-Actin (loading control; Sigma, St. Louis, MO, USA; catalog: A5441)

Suramin (Sigma, St. Louis, MO, USA; catalog: S2671)

iNOS (BD Pharmingen, San Diego, CA, USA; catalog: 610600)

COX-2 (Cayman Chemical, Ann Arbor, MI, USA; catalog: 160126)

NF-κB (Merck Millipore, Massachusetts, USA; catalog: MAB3026)

Ym-1 (Abcam, Biorbyt, Cambridge, UK; catalog: ab192029)

Arg-1 (Proteintech, Chicago, USA; catalog: 16001-1-AP)

PTP1B (Proteintech, Chicago, USA; catalog: 11334-1-AP)

Phospho-eukaryotic initiation factor 2 (p-eIF2) (antibodies-online; catalog: ABIN2776588)

Binding immunoglobulin protein (GRP-78) (Proteintech, Chicago, USA; catalog: 11587-1-AP)

Phospho-extracellular signal-regulated kinase (p-ERK) (Cell Signaling Technology, Danvers, MA, USA, Thr202/204; catalog: 9101)

Caspase-3 (IMGENEX, San Diego, CA, USA; catalog: Img-144A)

Tyrosine hydroxylase (Merck Millipore, Massachusetts, USA; catalog: MAB3018)

Feng C.W., Chen N.F., Chan T.F, & Chen W.F. (2020). Therapeutic Role of Protein Tyrosine Phosphatase 1B in Parkinson's Disease via Antineuroinflammation and Neuroprotection In Vitro and In Vivo. Parkinson's Disease, 2020, 8814236.

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Rat PCC PC12 Cell Culture and Analysis

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Rat PCC PC12 cells were acquired from American Type Culture Collection ATCC (Manassas, VA, USA) and cultured in our laboratory. Dulbecco's Modifi ed Eagle Medium (DMEM) was acquired from Mediatech (Herndon, VA, USA). Fetal bovine serum (FBS) and 6-OHDA (6-Hydroxydopamine) were acquired from Sigma-Aldrich (St Louis, MO, USA). All primary antibodies (antiβ-actin, anti-pAKT, anti-PTEN, anti-AKT) were acquired from Cell Signaling Technology (Boston, MA, USA).

Li L., Xu J., Wu M, & Hu J.M. (2018). Protective role of microRNA-221 in Parkinson's disease. Bratislavske lekarske listy, 119(1).

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Neuroprotective Potential of A5+ in Neuroblastoma Cells

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Murine neuroblastoma cell lines, N1E-115, were purchased from the American Type Culture Collection (ATCC, Manassas, VA, USA) and culture in Dulbecco’s Modified Eagle Medium (DMEM) (Gibco, Thermo Fisher Scientific, Waltham, MA, USA), supplemented with 10% heat-inactivated Fetal Bovine Serum (FBS) (Corning, NY, USA) and 100 U/mL penicillin/streptomycin (Thermo Scientific, Waltham, MA, USA). Cells were maintained at 37 °C in humidified air containing 5% CO₂.
A5⁺ was provided by SirtLife srl, Rome, Italy. A5⁺ 10 µg is composed of ellagic acid (20%), polydatin (98%), pterostilbene (20%), honokiol (20%), mixed with recommended doses of zinc, selenium, and chromium. The polydatin and A5⁺ were dissolved in DMSO (dimethyl sulfoxide, Sigma Aldrich, Milan, Italy) at 1 mg/mL [22 (link)]. In order to establish the concentration of A5⁺ able to promote beneficial effect without exerting neurotoxic effect, N1E115 cells were treated with different concentrations (10, 50, 100 and 500 μM) for 48 h.
Neurotoxicity was induced by treating cells with different concentration (10, 30, 50, 75 and 100 μM) of 6-hydroxydopamine (6-OHDA) (Sigma Aldrich, Saint Louis, MO, USA) for 24 h.
Moreover, a combined treatment has been given: cells were pre-treated with A5⁺ for 48 h and then 6-OHDA was added in combination with A5⁺ for further 24 h.

Pacifici F., Salimei C., Pastore D., Malatesta G., Ricordi C., Donadel G., Bellia A., Rovella V., Tafani M., Garaci E., Tesauro M., Lauro D., Di Daniele N, & Della-Morte D. (2022). The Protective Effect of a Unique Mix of Polyphenols and Micronutrients against Neurodegeneration Induced by an In Vitro Model of Parkinson’s Disease. International Journal of Molecular Sciences, 23(6), 3110.

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Neuroprotective Effects of SB Against 6-OHDA

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Neuroprotection assays were performed using alamarBlue^® reagent (Invitrogen, MA, USA). Briefly, SH-SY5Y cells were pretreated with SB for 1 h before adding 20 μM 6-hydroxydopamine (6-OHDA; Sigma, St. Louis, MO, USA) for 18 h. Cells were treated with LY294002 inhibitor (Sigma, St. Louis, MO, USA) or ZnPP (R&D Systems Inc., Minneapolis, MN, USA) prior to SB treatment for 1 h. Next, 10 μL alamarBlue^® was added after exposure to 6-OHDA, and optical density (OD) was measured with an ELISA reader. Relative protection (percent) was calculated as 100 × ((OD of the 6-OHDA + SB-treated cells − OD of the 6-OHDA-treated cells)/(OD of the control cells − OD of the 6-OHDA-treated cells)) as described previously [72 (link)].

Feng C.W., Chen N.F., Wen Z.H., Yang W.Y., Kuo H.M., Sung P.J., Su J.H., Cheng S.Y, & Chen W.F. (2019). In Vitro and In Vivo Neuroprotective Effects of Stellettin B Through Anti-Apoptosis and the Nrf2/HO-1 Pathway. Marine Drugs, 17(6), 315.

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Cell Culture and Animal Study Reagents

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For cell culture, fetal bovine serum (FBS) was purchased from Hyclone (Piscataway, NY, U.S.A.). Horse serum, penicillin, streptomycin, and MTT were purchased from Invitrogen (Carlsbad, CA, U.S.A.). Dulbecco’s modified Eagle’s medium (DMEM), L-glutamine, and LPS were purchased from Sigma-Aldrich Co. (St. Louis, MO, U.S.A.). For the animal study, 6-hydroxydopamine (6-OHDA) was purchased from Sigma-Aldrich Co. and diluted in saline containing 0.02% ascorbic acid. Desipramine (25 mg/kg) and 3,4-dihydroxyphenyl-L-alanine (L-DOPA, 20 mg/kg) were purchased from Sigma-Aldrich Co. and dissolved in saline. d-Amphetamine (d-AMPH) was purchased from USP (Rockville, MD, U.S.A.) and dissolved in saline. Compound 2 was dissolved in saline containing 0.25% ethanol.

Han B.S., Kim K.S., Kim Y.J., Jung H.Y., Kang Y.M., Lee K.S., Sohn M.J., Kim C.H., Kim K.S, & Kim W.G. (2016). Daphnane Diterpenes from Daphne genkwa Activate Nurr1 and Have a Neuroprotective Effect in an Animal Model of Parkinson’s Disease. Journal of natural products, 79(6), 1604-1609.

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Viral Injections and 6OHDA in Mice

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6–10-week-old mice were anesthetized and placed in a stereotaxic frame, and a 33-gauge needle (Plastics One) was inserted for viral injections into cortex (AP +0.5/1.5, ML +/−2.0, DV −1.0 from dura) or thalamus (AP −2.3, ML +/−0.5, DV −3.25 from dura). A volume of 0.75 μl/site of virus was injected at a rate of 0.2 μl/min. Two weeks later, the scalp was reopened and 1 μl of 5 μg/μl 6-hydroxydopamine (6OHDA; Sigma-Aldrich) was injected unilaterally into the medial forebrain bundle (MFB; AP −1.0, ML −1.0, DV −4.9 from dura) or bilaterally into the DLS (AP +0.8, ML +/− 2.2, DV −2.5 from dura).

Parker P.R., Lalive A.L, & Kreitzer A.C. (2016). Pathway-Specific Remodeling of Thalamostriatal Synapses in Parkinsonian Mice. Neuron, 89(4), 734-740.

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6-OHDA Stereotaxic Lesioning Protocol

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Animals were anaesthetized with a mixture of ketamine (8.7 mg/ml; Mavlab, Slack Creek, QLD) and xylazil (2 mg/ml; Troy Laboratories Pty Ltd, Smithfield, Australia) and placed in a stereotaxic apparatus (Kopf Instruments, Tujunga, California). Mice were then injected with 2 x 2 μl of 3 mg/mL (total 12 μg) of 6-hydroxydopamine (6-OHDA; Sigma Aldrich, Australia) in 0.02% ascorbic acid (sham) in the right sensorimotor striatum at the following coordinates: AP +1.0, ML +2.1, DV -2.9 and AP +0.3, ML +2.3, DV -2.9, relative to bregma and the dural surface as previously described [8 ]. 6-OHDA (or 0.02% ascorbic acid control) was injected at a rate of 0.5 μl/minute and the syringe left in place for 2 minutes after each injection to allow for complete diffusion.

Stayte S., Rentsch P., Li K.M, & Vissel B. (2015). Activin A Protects Midbrain Neurons in the 6-Hydroxydopamine Mouse Model of Parkinson’s Disease. PLoS ONE, 10(4), e0124325.

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Induction of Adipose Tissue Denervation in Obese Mice

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Diet-induced obese (DIO) mice were anesthetized with isoflurane and then the bilateral incisions were made approximately 0.5 cm. 6-hydroxydopamine (6-OHDA, Sigma-Aldrich, USA) was dissolved in PBS containing 1% ascorbic acid and subsequently injected into the bilateral subcutaneous fat pads (2 μL for per injection, 24 μL per pad), according to previously described 28 (link). The same volume of vehicles was injected into control mice. The bilateral incisions were closed as described above. The VSG and Sham mice were immediately prepared for surgery.

Jin Z., Meng W., Xiao T., Deng J., Wang J., Wen J., Chen K., Wang L., Liu J., Li Q., He J., Wang Z., Liu W, & Liu F. (2023). Vertical sleeve gastrectomy-derived gut metabolite licoricidin activates beige fat thermogenesis to combat obesity. Theranostics, 13(9), 3103-3116.

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Denervation and Cold Exposure in Mice

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7-week-old male C57BL/6 mice were anesthetized using constant flow isoflurane inhalation and a small incision was made in the abdominal skin. One of two inguinal fat (subcutaneous WAT (sWAT)) pads was denervated and the other pad, used as the control, was injected with the vehicle. 6-hydroxydopamine (6-OHDA, Sigma) was dissolved in PBS containing 1% ascorbic acid (9 mg/ml), and injected into the denervated pad using a 25 μl Hamilton syringe (2 μl for each injection, total 24 μl/pad) within 10 minutes. The same volume of vehicle, PBS containing 1% ascorbic acid was injected into the control fat pad. The needle was kept in place for 10 seconds after each injection to minimize reflux from the site of injection. The skin incision was closed with stainless steel wound clips and covered with nfz puffer to protect the wound from infection after injection. The mice were then administered with 1 mg/kg of buprenorphine by intraperitoneal injection. Two weeks post denervation, half of operated mice were individually housed in cages with free access to food and water continuously and exposed to cold (6 °C) for 2 days and all were then euthanized. Inguinal fat was isolated for flow cytometry analysis.

Ding X., Luo Y., Zhang X., Zheng H., Yang X., Yang X, & Liu M. (2016). IL-33-driven ILC2/eosinophil Axis in Fat Is Induced by Sympathetic Tone and Suppressed by Obesity. The Journal of endocrinology, 231(1), 35-48.

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Neuroinflammation Modulation Protocol

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EA (purity > 95%), LPS (Escherichia coli O111:B4), 6-hydroxydopamine (6-OHDA), and apomorphine hydrochloride were obtained from Sigma-Aldrich (St. Louis, CA, USA). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-1β, and IL-18 were bought from Elabscience Biotechnology Co., Ltd. (Wuhan, China). The MTT assay kit was from Beijing Solarbio Science and Technology Co., Ltd. (Beijing, China). Small interfering RNA (siRNA) against NLRP3 was purchased from GenePharma (Shanghai, China). Anti-CR3 complement receptor (OX-42 Catalog No. Ab1211) and tyrosine hydroxylase (TH, Catalog No. Ab113) antibodies were bought from Abcam (Cambridge, MA, USA). Anti-caspase-1 (Catalog No. 22915-1-AP), ionized calcium-binding adapter molecule-1 (Iba-1, Catalog No. 10904-1-AP), β-actin (Catalog No. 20536-1-AP), TNF-α (Catalog No. 17590-1-AP), IL-1β (Catalog No. 66737-1-Ig), IL-18 (Catalog No. 10663-1-AP), rabbit IgG (Catalog No. SA00001-2), and mouse IgG (Catalog No. SA00001-1) antibodies were purchased from the Proteintech Group (Chicago, IL, USA). Anti-NLRP3 (Catalog No. orb101128) antibody was purchased from Biorbyt (Cambridge, United Kingdom).

He X.M., Zhou Y.Z., Sheng S., Li J.J., Wang G.Q, & Zhang F. (2020). Ellagic Acid Protects Dopamine Neurons via Inhibition of NLRP3 Inflammasome Activation in Microglia. Oxidative Medicine and Cellular Longevity, 2020, 2963540.

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