Acitretin

Manufactured by Merck Group

Sourced in United States

Acitretin is a prescription pharmaceutical product manufactured by Merck Group. It is a lab equipment used for analytical and research purposes. Acitretin is a synthetic retinoid compound.

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13 protocols using acitretin

Acitretin Administration for ZIKV Treatment

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After 6 d of treatment with 1 mg/mouse Depo-provera, 20 µl of 2 mM acitretin (Sigma-Aldrich) in DMSO (Hsia et al., 2008 (link)) was administered into the vaginal cavity at the indicated times either before or after i.vag. ZIKV inoculation. All mice were anesthetized in an isoflurane chamber before administering the drug.

Khan S., Woodruff E.M., Trapecar M., Fontaine K.A., Ezaki A., Borbet T.C., Ott M, & Sanjabi S. (2016). Dampened antiviral immunity to intravaginal exposure to RNA viral pathogens allows enhanced viral replication. The Journal of Experimental Medicine, 213(13), 2913-2929.

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Latency Reactivation Screening Protocol

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50,000 J-Lat 10.6 and JNLGFP cells were aliquoted into 96-well plates in 200 μL medium. Each of the following drugs were added to the cells in triplicate at the indicated concentrations: BMH-21 (MedKoo, 406586), RA190 (Calbiochem, 5.30341.0001), ERW1041E (Sigma, 509522), ZDON (Calbiochem, 616467), ZDON (Calbiochem, 15227), tubacin (Sigma, SML0065), b-AP15 (Cayman chemical, 11324; AdooQ Bioscience, A15391; MedKoo, 406452), capzimin (Glix labs, GLXC-09966), thiolutin (MedKoo, 525293; Cayman chemical, 11350), WP1130 (Cayman chemical, 15227), P005091 (MedKoo, 406577; Cayman chemical, 15224), IU1 (AdooQ, A13209; Cayman chemical, 10617), TCID (Cayman chemical, 16353; Sigma, SML1402), acitretin (Sigma, 44707), prostratin (Sigma, P0077), ingenol (Sigma, SML1318), vorinostat (Sigma, SML0061), JQ1 (Sigma, SML1524), I-BET151 (Tocris, 4650), 5-azacytidine (Sigma, A3656), romidepsin (Sigma, SML1175), PR619 (AdooQ Bioscience, A13190). For control groups, 0.1% DMSO was used. After 48 or 72 h of treatment, GFP expression in the cells were measured by flow cytometry. Cell viabilities were determined by forward scatter vs. side scatter gating using untreated cells as the control. The data were analyzed with FlowJo software and plotted as bar graphs.

Rathore A., Iketani S., Wang P., Jia M., Sahi V, & Ho D.D. (2020). CRISPR-based gene knockout screens reveal deubiquitinases involved in HIV-1 latency in two Jurkat cell models. Scientific Reports, 10, 5350.

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Retinoid Compound Quantification Protocol

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All-trans-RA (atRA), 9-cis-RA (9cRA), 13-cis-RA (13cRA), and acitretin were purchased from Sigma-Aldrich (St. Louis, MO). 9,13-di-cis-RA (9,13dcRA) was prepared as described.^{33 (link)} acitretin was used as the internal standard. Optima LC/MS grade water (H₂O), acetonitrile (ACN), and formic acid (FA) were purchased from Fisher Scientific (Pittsburgh, PA).

Jones J.W., Pierzchalski K., Yu J, & Kane M.A. (2015). Use of Fast HPLC Multiple Reaction Monitoring Cubed for Endogenous Retinoic Acid Quantification in Complex Matrices. Analytical chemistry, 87(6), 3222-3230.

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ADAM10 activation and inhibition protocol

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To activate ADAM10, cells were incubated in Opti-MEM containing GlutaMAX (Life Technologies) containing either 20 μm carbachol (Sigma) for 24 h or 20 μm acitretin (Sigma) for 48 h. To inhibit ADAM10, cells were incubated with 10 μm GI254023X (Tocris), a selective ADAM10 inhibitor, for either 24 or 48 h in Opti-MEM. DMSO only–treated cells were used for comparison with treated cells.

Jarosz-Griffiths H.H., Corbett N.J., Rowland H.A., Fisher K., Jones A.C., Baron J., Howell G.J., Cowley S.A., Chintawar S., Cader M.Z., Kellett K.A, & Hooper N.M. (2019). Proteolytic shedding of the prion protein via activation of metallopeptidase ADAM10 reduces cellular binding and toxicity of amyloid-β oligomers. The Journal of Biological Chemistry, 294(17), 7085-7097.

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Measuring Long-Lived Protein Degradation

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NB4 cells were seeded at a density of 0.1 × 10⁶/ml and treated with vehicle, 1 µM ATRA (Sigma-Aldrich, Buchs, Switzerland) or 1 µM acitretin (Sigma-Aldrich, Buchs, Switzerland) for two days before the addition of 0.2µCi of ¹⁴C-Valin/ml (L-(U-14-C)Valine, Code CFB.75, Amersham). Cells were incubated for another two days before the long-lived protein degradation assays were performed. Cells were washed and incubated for 1.5 h in the presence of 10 mM non-radioactive L-Valin before the 5 h chasing phase was started. Protein precipitations were carried using trichloracetic acid (at 10%) and phosphotunctic acid (at 1%). After incubation at 4 °C for 30 minutes samples were centrifuged at 600 rcf for 15 minutes. Radioactivity in the supernatant and the pellet fractions was determined using a scintillation counter (PerkinElmer). Proteolysis was calculated as the percentage of radioactivity in the supernatant.

Schläfli A.M., Isakson P., Garattini E., Simonsen A, & Tschan M.P. (2017). The autophagy scaffold protein ALFY is critical for the granulocytic differentiation of AML cells. Scientific Reports, 7, 12980.

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Formulation of PLGA-based Implants

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50:50 acid end-capped PLGA 503H (24–38 kDa), was purchased from Evonik, 4HPR was generously supplied by Merck Co, and acitretin was used as an internal standard (analytical grade, Sigma-Aldrich). Excipients used were sodium deoxycholate, (NaDC, 99% pure, Acros), polyvinylpyrrolidone (PVP K30, 40 kDa, BASF), hydroxypropyl methylcellulose K4M (HPMC K4M, Dow Chemical, Midland, MI), and β-CD (Sigma-Aldrich). Calcium deoxycholate (CaDC) was synthesized according to literature methods described in Supplemental information. Water-soluble implants were composed of polyvinyl alcohol (PVA, 9–10 kDa, 80% hydrolyzed, Sigma-Aldrich) and _D-sucrose (Sigma-Aldrich). All other materials were reagent grade or better including: MgCO₃, acetone, ethanol (EtOH), tetrahydrofuran (THF), and Tween 80. Solvents for UPLC-UV and MS analysis were HPLC or MS grade including acetonitrile (ACN), methanol (MeOH), double distilled water (ddH₂O), phosphoric acid (H₃PO₄), ammonium formate (NH₄COO) and formic acid (HCOOH). Silicon tubing (0.8 mm i.d.,) was purchased from BioRad Laboratories. Ethylenediaminetetraacetic acid (EDTA, analytical standard, LECO, St. Joseph, MI) and Bovine Serum Albumin (BSA, 99% pure, heat shock fraction, Sigma-Aldrich) were used as standards for nitrogen analysis.

Nieto K., Pei P., Wang D., Mallery S.R, & Schwendeman S.P. (2017). In vivo controlled release of fenretinide from long-acting release depots for chemoprevention of oral squamous cell carcinoma recurrence. International journal of pharmaceutics, 538(1-2), 48-56.

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Sustained Retinoid Delivery from PLLA

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ATRA (CAS 302-79-4, purity ≥ 98%), acitretin (CAS 55079-83-9, purity ≥ 98%), fluorescein isothiocyanate isomer I (FITC) (CAS 3326-32-7, purity ≥ 90%), and corn oil (CAS 8001-30-7, d: 0.9 g/mL) were purchased from Sigma-Aldrich (MO, USA). Ester-terminated PLLA (Mw 50,000 g/mol) was obtained from Jinan Daigang Biomaterial Co., Ltd. (Jinan, China). CO₂ (purity > 99.9%, v/v) was supplied by Xiamen Rihong Co., Ltd. (Xiamen, China). ATRA slow-releasing pellets were purchased from Innovative Research of America (Florida, USA). All other chemical reagents were of analytical purity.

Yang D., Luo W., Wang J., Zheng M., Liao X.H., Zhang N., Lu W., Wang L., Chen A.Z., Wu W.G., Liu H., Wang S.B., Zhou X.Z, & Lu K.P. (2017). A novel controlled release formulation of the Pin1 inhibitor ATRA to improve liver cancer therapy by simultaneously blocking multiple cancer pathways. Journal of controlled release : official journal of the Controlled Release Society, 269, 405-422.

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Quantitative ATRA Detection in Eosinophils and DCs

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A LC(liquid chromatography)/MS/MS-based method was established for quantitative detection of ATRA in cell cultures referring to a previous report[17 (link)]. Briefly, 10⁵ CD11b⁺ Siglec-F⁺ CCR3⁺ LP or PB eosinophils or CD11c⁺MHC-II⁺CD103⁺ LP DC was mixed with 1 ml 0.025 M KOH-ethanol, and then added internal standard. The mixture was added with 10 ml hexane for vortex, followed by a short centrifugation for stratification. The lower aqueous phase was collected to mix with 60 μl of 4 M HCl and then added with hexane as above. After centrifugation, the hexane phase was collected to remove the solvent with a gentle nitrogen stream. The residue was dissolved in 100 μl acetonitrile for measurement of ATRA using a triple-quadrupole mass spectrometer (Agilent Technologies, USA). In the detection, acitretin (Sigma), an analogue of ATRA, was used as the internal reference[18 (link)].

Chen H.H., Sun A.H., Ojcius D.M., Hu W.L., Ge Y.M., Lin X., Li L.J., Pan J.P, & Yan J. (2015). Eosinophils from Murine Lamina Propria Induce Differentiation of Naïve T Cells into Regulatory T Cells via TGF-β1 and Retinoic Acid. PLoS ONE, 10(11), e0142881.

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ATRA-Loaded Nanoparticle Cancer Therapy

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ATRA, acitretin, fluorescein isothiocyanate isomer I (FITC) and corn oil were acquired from Sigma-Aldrich (Missouri, USA). The 21-day ATRA sustained-release pellet was obtained from Innovative Research of America (Florida, USA). PLA–PEG–PLA triblock polymers (Mw 110 kDa, PEG 9%) were obtained from Jinan Daigang Biomaterial (Shandong, China). Cell lysis buffer for IP, BCA protein assay kit, protease inhibitor cocktail, 4′, 6-diamidino-2-phenylindole (DAPI) kit, hematoxylin and eosin (H&E) kit, crystal violet staining solution and citrate–EDTA antigen retrieval solution were procured from Beyotime Biotechnology (Shanghai, China).
Hepatocellular carcinoma cell lines were provided by the Cell Bank (Shanghai, China). Cells were cultured with Dulbecco’s Modified Eagle Medium (DMEM; ThermoFisher, USA) containing 10% fetal bovine serum (FBS; PAN, Germany) and 1% penicillin–streptomycin solution (HyClone, USA) at 37°C under 5% CO₂ in a humidified incubator. Male BALB/c nu/nu mice were obtained from Shanghai Laboratory Animal Center (SLAC, Shanghai, China). The animal care and experimental protocols were performed in accordance with and approved by the Experimental Animal Ethics Committee of Fujian Medical University.

Zhang F., Zhang A., Xie Y., Wen H., Kankala R.K., Huang J., Zhang A., Wang Q., Chen B., Dong H., Guo Z., Chen A, & Yang D. (2023). Nanocarrier of Pin1 inhibitor based on supercritical fluid technology inhibits cancer metastasis by blocking multiple signaling pathways. Regenerative Biomaterials, 10, rbad014.

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Culturing T Cells and HIV Infection

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All T cells and T cell lines were cultured in RPMI 1600 supplemented with penicillin (50 U/ml), streptomycin (50 µg/ml), L-glutamine (2 mM) (all from Life Technologies, Grand Island, NY), and 10% heat-inactivated fetal calf serum (FCS) (from Sigma, St. Louis, MO). TZM-bl cells were cultured in DMEM supplemented with the same additives as above. During cell culture post treatment or activation, 1 µM indinavir (IDV), 10 µM nevirapine (NVP), and 600 nM raltegravir (RAL) (NIH AIDS Reagent Program) were included to prevent spreading HIV infection. Working stocks of acitretin and SAHA were dissolved in DMSO (all from Sigma, St. Louis, MO). anti-CD3 and anti-CD28 antibodies beads (CD3/28) (Life Technologies, Grand Island, NY) were used at 1 bead per cell with human IL2 (Chiron, Emeryville, CA) at 10U/ml.

Li P., Kaiser P., Lampiris H.W., Kim P., Yukl S.A., Havlir D.V., Greene W.C, & Wong J.K. (2016). Stimulating the RIG-I pathway to kill cells in the latent HIV reservoir following viral reactivation. Nature medicine, 22(7), 807-811.

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