Tunicamycin

Manufactured by Apexbio

Sourced in United States

Tunicamycin is a laboratory reagent used in cell biology research. It functions as an inhibitor of N-linked glycosylation, a process involved in the synthesis of glycoproteins. Tunicamycin is commonly employed in studies to investigate the role of glycosylation in various cellular processes.

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Lab products found in correlation

Oligomycin, by Merck Group (1 mentions) Lamin b, by Proteintech (1 mentions) Phospho gsk3β, by Cell Signaling Technology (1 mentions) Swainsonine s9263, by Merck Group (1 mentions) Unc 2250, by Selleck Chemicals (1 mentions) Phospho mertk, by Abcam (1 mentions) Pdhk1, by Cell Signaling Technology (1 mentions) Mertk, by Abcam (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Phospho akt ser473, by Cell Signaling Technology (1 mentions) D glucose, by Merck Group (1 mentions) Diethylnitrosamine, by Merck Group (1 mentions) β actin, by Proteintech (1 mentions) Sybr green, by Bio-Rad (1 mentions) Gapdh, by Abmart (1 mentions) Rpmi 1640, by Solarbio (1 mentions) Hp β cd, by BBI Lifesciences (1 mentions) Cycloheximide (chx), by Apexbio (1 mentions) Bortezomib ps 341, by Selleck Chemicals (1 mentions) Actinomycin d, by Apexbio (1 mentions)

8 protocols using tunicamycin

Evaluating Metabolic Reprogramming Mechanisms

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Diethylnitrosamine (DEN, 73861), CCl₄ (1601168), Oligomycin (O4876), Carbonyl cyanide 4-(trifluoromethoxy) phenylhydrazone (FCCP, C2920), antimycin A (A8674), rotenone (R8875), 2-deoxy-d-glucose (2-DG, D8375), PNGase F (G5166) and Swainsonine (S9263) and D-(+)-Glucose (G7021) were obtained from Sigma (St. Louis, MO, USA). SYBR green was purchased from Bio-Rad (Hercules, CA, USA). Horseradish peroxidase (HRP)-conjugated anti-rabbit, anti-mouse immunoglobulin G were obtained from Beyotime (Haimen, Jiangsu, China). Glucose deficient DMEM medium and Trizol Reagent were obtained from Life Technologies (Carlsbad, CA, USA). Tunicamycin (B7417) was obtained from ApexBio Technology (Houston, TX, USA). UNC-2250 (S7342) was purchased from Selleck Chemicals (Houston, TX, USA). The antibodies used in this study are: MerTK (abcam; ab52968), Phospho-MerTK (abcam; ab14921), Akt (Cell Signaling Technology; 4691), Phospho-Akt (Ser473) (Cell Signaling Technology; 4060), Phospho-GSK3β (Cell Signaling Technology; 5558S), PGK1 (ZN; 501965), LDHA (ZN; 501146), PFKM (ZN; 505477), PKM2 (ZN; 505477), PDHK1 (Cell Signaling Technology; 3820S), Lamin B (Proteintech; 12987-1-AP), GAPDH (Abmart; M20028) and β-Actin (Proteintech; 60008-1-1 g).

Liu Y., Lan L., Li Y., Lu J., He L., Deng Y., Fei M., Lu J.W., Shangguan F., Lu J.P., Wang J., Wu L., Huang K, & Lu B. (2022). N-glycosylation stabilizes MerTK and promotes hepatocellular carcinoma tumor growth. Redox Biology, 54, 102366.

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Cholesterol Depletion and ER Stress in Breast Cancer

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MDA-MB-231 and BT-549 cells were obtained from the Cell Bank of Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China). The cells were maintained in a basal medium (RPMI-1640, Solarbio Science & Technology Co., Ltd., Beijing, China) containing 10% foetal bovine serum (FBS). To deplete cholesterol, cells were treated with HP-β-CD (BBI life Sciences, Shanghai, China) for 48 h. The regulation of cholesterol efflux and synthesis was carried out by culturing cells in an experimental medium containing LXR-623 (agonist of LXRα, cat. no. HY-10629; MedChemExpress, Monmouth Junction, NJ, USA), simvastatin, or DMSO (<0.1%) for 24 h. To induce ER stress, cells were incubated with tunicamycin (APExBIO technology, Houston, TX, USA, cat. no. B7417) for 24 h.

Zhao Y., He L., Wang T., Zhu L, & Yan N. (2021). 2-Hydroxypropyl-β-cyclodextrin Regulates the Epithelial to Mesenchymal Transition in Breast Cancer Cells by Modulating Cholesterol Homeostasis and Endoplasmic Reticulum Stress. Metabolites, 11(8), 562.

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Modulation of Cellular Responses

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Tunicamycin, Cycloheximide and Actinomycin D were obtained from APExBio (B7414, A8244 and A4448). Recombinant human interferon γ and recombinant mouse interferon γ were obtained from Meilun Biotechnology co., Ltd. Dalian (MB5954 and MA0620). Bortezomib (PS-341) was obtained from Selleck (S1013).

Zhang J., Zhang G., Zhang W., Bai L., Wang L., Li T., Yan L., Xu Y., Chen D., Gao W., Gao C., Chen C., Ren M., Jiao Y., Qin H., Sun Y., Zhi L., Qi Y., Zhao J., Liu Q., Liu H, & Wang Y. (2022). Loss of RBMS1 promotes anti-tumor immunity through enabling PD-L1 checkpoint blockade in triple-negative breast cancer. Cell Death and Differentiation, 29(11), 2247-2261.

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Tunicamycin-Induced ER Stress Model

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For the ER stress model, mice were intraperitoneally injected with tunicamycin (APExBIO) dissolved in dimethyl sulfoxide (DMSO) and diluted in sterile 150 mM dextrose at a dose of 0.5 μg/g body mass at ZT8, and tissues were sampled 24 h later.

Huang K., Zhang T., Zhang W., Gu Y., Yu P., Sun L., Liu Z., Wang T, & Xu Y. (2023). Multigenerational mistimed feeding drives circadian reprogramming with an impaired unfolded protein response. Frontiers in Endocrinology, 14, 1157165.

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Culturing Human Lung Cancer Cell Lines

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Human lung cancer cell lines H358, H1299 and H2170 were obtained from ATCC (American Type of Cell Collection, Manassas, VA, USA). Non-immortalized MEF cells were cultured and maintained by our laboratory. Cells were cultured in RPMI 1640 medium (for H358 and H2170) or DMEM medium (for H1299 and MEF), supplemented with 10% fetal bovine serum (extra 1% non-essential amino acids for MEF) and incubated in a 5% CO₂ humidified chamber at 37 °C. MLN4924 (#B1036) and tunicamycin (#B7417) was purchased from ApexBio. Chloroquine (CQ, #C6628), chlorhexidine (CHX, #C7698) and N-acetyl-l-cysteine (NAC, #A7250) were purchased from Sigma. MG132 (#HY-13259) was purchased from MedChem Express.

Li Y., Wang C., Xu T., Pan P., Yu Q., Xu L., Xiong X., Hou T., Cui S, & Sun Y. (2021). Discovery of a small molecule inhibitor of cullin neddylation that triggers ER stress to induce autophagy. Acta Pharmaceutica Sinica. B, 11(11), 3567-3584.

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Autophagy Regulation in Cancer Cells

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Anti-ATG5 (C-1; cat. no. sc-133158; 1:500), anti-MAP-LC3-β (G-9; cat. no. sc-376404; 1:500) and anti-sequesteome 1 (SQSTM1; D-3; cat. no. SC-28359; 1:500) primary antibodies were purchased from Santa Cruz Biotechnology, Inc. Anti-β-actin (ACTB; cat. no. 100166-MM10; 1:500) primary antibody was obtained from Sino Biological, Inc., and the anti-clathrin primary antibody was obtained from Covance, Inc. The PCR primers were synthesized by Sangon Biotech Co., Ltd. Atorvastatin calcium was purchased from Sigma-Aldrich (Merck KGaA). Rapamycin and bortezomib were purchased from LC Laboratories. Chloroquine was obtained from Cell Signaling Technology, Inc. EGF was obtained from PeproTech, Inc., and docetaxel from Absin Bioscience Inc. Valproic acid (VPA), brefeldin A, tunicamycin and thapsigargin were obtained from APeXBIO Technology LLC. The Annexin V-FITC apoptosis detection kit (cat. no. AD10) was purchased from Dojindo Molecular Technologies, Inc. All cancer cell lines were purchased from the American Type Culture Collection.

Peng K., Sun A., Zhu J., Gao J., Li Y., Shao G., Yang W, & Lin Q. (2021). Restoration of the ATG5-dependent autophagy sensitizes DU145 prostate cancer cells to chemotherapeutic drugs. Oncology Letters, 22(3), 638.

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Tunicamycin and MFG-E8 Treatment Assay

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The AR42J cells were treated with 0.5 or 1 μM tunicamycin (B7417, APExBIO, Houston, United States) with or without 20 or 100 ng/ml MFG-E8 (2805-MF, RD System, Inc. Minnesota, United States) for 24 h. Protein homogenate was extracted for subsequent detection.

Ren Y., Liu W., Zhang J., Bi J., Fan M., Lv Y., Wu Z., Zhang Y, & Wu R. (2022). MFG-E8 Maintains Cellular Homeostasis by Suppressing Endoplasmic Reticulum Stress in Pancreatic Exocrine Acinar Cells. Frontiers in Cell and Developmental Biology, 9, 803876.

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Dopamine Antagonists Screen for eIF2α Phosphorylation

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A549 cells were treated with 27 FDA-approved dopamine receptor antagonists (from Selleck Chemicals, Houston, TX, USA) at a concentration of 15 μM for 12 h. Then, cells were collected to obtain protein and P-eIF2α level was detected. Twenty-seven drugs were as follows: Amisulpride (S1280), Paliperidone (S1724), Quetiapine Fumarate (S1763), Chlorprothixene (S1771), Tetrabenazine (Xenazine) (S1789), Haloperidol (S1920), Pramipexole 2HCl Monohydrate (S2011), Levosulpiride (S2104), Amantadine HCl (S2451), Pramipexole (S2460), Domperidone (S2461), Dopamine HCl (S2529), Benztropine mesylate (S3163), (+, −)-Octopamine HCl (S3188), Ropinirole HCl (S3189), Trifluoperazine 2HCl (S3201), Pergolide Mesylate (S4000), Droperidol (S4096), Penfluridol (S4151), Azaperone (S4219), Rotigotine (S4274), Metoclopramide HCl (S4289), Fluphenazine dihydrochloride (S4569), Fenoldopam mesylate (S4618), Prochlorperazine dimaleate salt (S4631), Brexpiprazole (S4639), Sulpiride (S4655). Tunicamycin was from APExBIO (Houston, USA, #B7417).

Mao Y., Wang Z., Yao C., Zeng Q., Cheng W., Zhang S., Chen S, & Sheng C. (2022). The Food and Drug Administration-approved antipsychotic drug trifluoperazine, a calmodulin antagonist, inhibits viral replication through PERK-eIF2α axis. Frontiers in Microbiology, 13, 979904.

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