Cycloheximide 01810

Manufactured by Merck Group

Sourced in United States

Cycloheximide (01810) is a laboratory reagent commonly used as a protein synthesis inhibitor. It functions by blocking the translocation step in eukaryotic protein synthesis, thereby inhibiting the growth of eukaryotic organisms.

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Lab products found in correlation

Anti myc sc 40, by Santa Cruz Biotechnology (1 mentions) Anti ha 3724, by Cell Signaling Technology (1 mentions) Mg132, by Merck Group (1 mentions) Torin1, by Bio-Techne (1 mentions) Chloroquine c6628, by Merck Group (1 mentions) Bafilomycin a1, by Fujifilm (1 mentions) Lysotracker red, by Thermo Fisher Scientific (1 mentions) Phospho stat3 9131, by Cell Signaling Technology (1 mentions) β actin sc 47778, by Santa Cruz Biotechnology (1 mentions) Pdgf bb, by R&D Systems (1 mentions) Pmturquoise2 er, by Addgene (1 mentions) Pmturquoise2 peroxi, by Addgene (1 mentions) Pmturquoise2 golgi, by Addgene (1 mentions) Roxadustat, by Cayman Chemical (1 mentions) Pa5 29975, by Thermo Fisher Scientific (1 mentions) Cip p4978, by Merck Group (1 mentions) Ndufa9 459100, by Thermo Fisher Scientific (1 mentions) Pgc1α h 300, by Santa Cruz Biotechnology (1 mentions) Dimethyl sulfoxide d8418, by Merck Group (1 mentions) Gst b 14, by Santa Cruz Biotechnology (1 mentions)

6 protocols using cycloheximide 01810

Immunodetection of UBE2O and Mxi1 Proteins

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Anti-UBE2O antibody (GTX119315) for immunohistochemical (IHC) analysis was purchased from GeneTex. Anti-UBE2O antibody (A301-873A) for immunoblotting was purchased from Bethyl Laboratories. Anti-Mxi1 antibody (HPA035319), anti-Flag antibody (F1804), and Cycloheximide (01810) were purchased from Sigma-Aldrich. Anti-Myc (sc-40) and anti-Mxi1 (sc-1042) antibodies were obtained from Santa Cruz Biotechnology. Anti-GAPDH (60004-1-Ig) antibody was obtained from Proteintech. Anti-HA (#3724) and anti-GST (#2624) antibodies were obtained from Cell Signaling Technology. Anti-ATM (A19650) and anti-phospho-ATM-Ser1981 (AP0008) antibodies were obtained from ABclonal. The proteasome inhibitor MG132 (474790) was purchased from Millipore. Arsenic trioxide (ATO) was a clinically available drug which was obtained from Beijing ShuangLu Pharmaceutical Co., Ltd., China (approval number: H20080664).

Huang Y., Yang X., Lu Y., Zhao Y., Meng R., Zhang S., Dong X., Xu S, & Wu G. (2020). UBE2O targets Mxi1 for ubiquitination and degradation to promote lung cancer progression and radioresistance. Cell Death and Differentiation, 28(2), 671-684.

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Autophagy Regulation: Antibody Toolkit

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Rabbit polyclonal antibody against EIF4EBP1/4E-BP1 (9542), RPS6KB1 (2217), and RPS6KB1 pThr389 (9234) were purchased from Cell Signaling. Guinea pig polyclonal and rabbit polyclonal anti-SQSTM1/p62 antibodies were purchased from Progen (GP62-C) and MBL (PM 045), respectively. Anti-GFP (NACALAI TESQUE, 04404) and anti-Multi Ubiquitin (MBL, D058-3) antibodies were purchased. Anti-LC3B⁴⁵ (link) antibody production was described previously. Horseradish peroxidase-conjugated anti-mouse (315035003) and anti-rabbit (111035144) IgG antibodies were purchased from Jackson ImmunoResearch, bafilomycin A₁ (02911643) and α-amanitin (23109059) were purchased from Wako, cycloheximide (01810) and chloroquine (C6628) were purchased from Sigma, and Torin1 was purchased from Tocris Biosciences (4247). LysoTracker Red (L7528) was purchased from Molecular Probes.

Sahani M.H., Itakura E, & Mizushima N. (2014). Expression of the autophagy substrate SQSTM1/p62 is restored during prolonged starvation depending on transcriptional upregulation and autophagy-derived amino acids. Autophagy, 10(3), 431-441.

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Molecular Signaling Pathway Analysis

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Vanillin (V1104), deferoxamine (D9533), MG132 (M7449), dimethyl sulfoxide (DMSO), protease inhibitor cocktail, and cycloheximide (01810) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against 4E-BP1 (9452), phospho-4E-BP1 (2855), eIF4E (9742), phospho-eIF4E (9741), phospho-S6 (4857), STAT3 (12640), and phospho-STAT3 (9131) were obtained from Cell Signaling Technology (Danvers, MA, USA). Antibodies against β-tubulin (sc-9104) and β-actin (sc-47778) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Anti-HIF-1α and anti-HIF-1β antibodies were kindly provided by Jong-Wan Park of Seoul National University, Seoul, Korea.

Park E.J., Lee Y.M., Oh T.I., Kim B.M., Lim B.O, & Lim J.H. (2017). Vanillin Suppresses Cell Motility by Inhibiting STAT3-Mediated HIF-1α mRNA Expression in Malignant Melanoma Cells. International Journal of Molecular Sciences, 18(3), 532.

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Isolation and Culture of Rat Vascular Smooth Muscle Cells

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Rat primary vascular smooth muscle cells (VSMCs) were cultured from rat thoracic aorta as described previously [29 ]. Rat thoracic aortas were removed and washed with PBS for 3 times. Briefly, aortic media layer was dissected, cut into pieces, and explanted onto a 6 well culture plate. Cells were growing at 37°C in a humidified atmosphere of 5% CO2 in DMEM supplemented with 10% FBS, penicillin and streptomycin for 2 weeks. VSMC phenotype were confirmed by expression of smooth muscle marker protein SM22α. PDGF-BB was obtained from R&D Systems (220-BB-010, Minneapolis, MN). Cycloheximide (01810) and pifithrin-α (P4359) were obtained from Sigma-Aldrich (St. Louis, MO, USA). p53 (2524S), pCDK1 (4539), CDK1 (9112), HA-tag (2367), Flag-tag (14793S), p-p53 (9284) antibodies were purchased from Cell Signaling (Danvers, MA, USA). GAPDH (G8795) antibody were obtained from Sigma-Aldrich (St. Louis, MO, USA). PCNA (sc-56), Caspase 3 (sc-56053), NOXA (sc-56169), p21 (sc-6246), p300 (sc-48343), MDM2 (sc-965) antibodies was purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Tang R., Mei X., Wang Y.C., Cui X.B., Zhang G., Li W, & Chen S.Y. (2019). LncRNA GAS5 regulates vascular smooth muscle cell cycle arrest and apoptosis via p53 pathway. Biochimica et biophysica acta. Molecular basis of disease, 1865(9), 2516-2525.

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Ubiquitination Compound Library for Drug Screening

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A ubiquitination compound library (L8600) for drug screening was obtained from TargetMol (Boston, MA, USA). VH298 (SML1896) and cycloheximide (01810) were purchased from Sigma-Aldrich (St. Louis, MO, USA). Roxadustat (15294) was obtained from Cayman Chemical Co. (Ann Arbor, MI, USA). VL285 (S0095) was obtained from Selleck Chemicals (Houston, TX, USA). Short interfering RNA (siRNA) targeting NBR1 (5′-GAAGAGGUAUCCAUCAACAUU-3′), p62/SQSTM1 (5′-GCAUUGAAGUUGAUAUCGAUUU-3′), HIF-1α (5’-CAAUCAAGAAGUUGCAUUA-3’), and scrambled siRNA (5′-CCUACGCCACCAAUUUCGU-3′) were synthesized by Genolution (Seoul, Republic of Korea). The subcellular localization vectors, pmTurquoise2-ER, pmTurquoise2-Golgi, pmTurquiose2-Mito, and pmTurquoise2-Peroxi, in which a cyan fluorescence protein (Turquoise) was fused with an ER, Golgi, mitochondria, or peroxisome-targeting sequence, respectively, were obtained from Addgene (36204, 36205, 36208, and 36203; deposited by Dorus Gadella). pcDNA3.1-HA (HA) was obtained from Addgene (128034; deposited by Oskar Laur). HA-tagged ubiquitin WT (HA-UB) was obtained from Addgene (17608; deposited by Ted Dawson).

Kim Y.H., Park N.Y., Jo D.S., Bae J.E., Kim J.B., Park K., Jeong K., Kim P., Yeom E, & Cho D.H. (2024). Inhibition of VHL by VH298 Accelerates Pexophagy by Activation of HIF-1α in HeLa Cells. Molecules, 29(2), 482.

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Cellular Protein Quantification and Validation

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Cycloheximide (01810) and calf-intestinal alkaline phosphatase (CIP, P4978) were purchased from Sigma. Dimethyl sulfoxide (D8418, Sigma-Aldrich, St Louis, MO, USA) was added to the cultures at 0.1% (v/v) as a solvent control. Specific antibodies against ATP5C1 (PA5-29975) and NDUFA9 (459100) were purchased from Invitrogen. Antibodies specific for ATP5A1 (51), SIRT3 (14.45), VDAC (B-6), SDHA (B-1), UQCRC2 (G-10), COX IV (D-20), PGC-1α (H-300), PGC-1β (E-9), NRF1 (H-285), NRF2 (C-20), Tfam (H-203), SIRT1 (H-300), PKCα (C-20), actin (I-19), V5 (H-9), Flag (D-8), His (AD1.1.10) and GST (B-14) were purchased from Santa Cruz Biotechnology (Dallas, TX, USA).

Kim Y.R., Kim J.S., Yun J.S., Kim S., Kim S.Y., Jang K, & Yang C.S. (2018). Toxoplasma gondii GRA8 induces ATP5A1–SIRT3-mediated mitochondrial metabolic resuscitation: a potential therapy for sepsis. Experimental & Molecular Medicine, 50(3), e464-.

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