Thapsigargin

Manufactured by R&D Systems

Sourced in United States

Thapsigargin is a natural product compound obtained from the plant Thapsia garganica. It functions as a specific and potent inhibitor of the sarco/endoplasmic reticulum Ca2+-ATPase (SERCA) pump, which plays a crucial role in regulating intracellular calcium homeostasis.

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Lab products found in correlation

Picrotoxin, by R&D Systems (8 mentions) Heparin, by R&D Systems (8 mentions) U73122, by R&D Systems (8 mentions) Chelerythrine, by R&D Systems (7 mentions) Kynurenic acid, by R&D Systems (7 mentions) U73343, by R&D Systems (6 mentions) Ml 133, by R&D Systems (6 mentions) Tertiapin q, by R&D Systems (5 mentions) Gdp β s, by R&D Systems (5 mentions) Bisindolylmaleimide 2 bis 2, by R&D Systems (5 mentions) Dioctanoyl phosphatidylinositol 4 5 bisphosphate dic8 pip2, by Echelon Biosciences (5 mentions) Sr49059, by R&D Systems (4 mentions) Ml 297, by R&D Systems (4 mentions) Tertiapin lq, by R&D Systems (3 mentions) Bapta, by R&D Systems (3 mentions) Bis 2, by R&D Systems (2 mentions) Capsazepine, by R&D Systems (2 mentions) Tunicamycin, by Merck Group (1 mentions) Mg132, by Merck Group (1 mentions) Etoposide, by Merck Group (1 mentions)

13 protocols using thapsigargin

Cellular Toxicity Screening Assay

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Aphidicolin, doxorubicin, rapamycin, chloroquine, trichostatin A, suberanilohydroxamic acid, etoposide, tunicamycin, MG132, nocodazole, and roscovitine were obtained from Sigma-Aldrich (St. Louis, MO). Thapsigargin (R&D Systems, Minneapolis, MN) was generously provided by Dr. Kelly Jordan-Sciutto. Aggregatibacter actinomycetemcomitans cytolethal distending toxin B subunit was obtained from Dr. Bruce Shenker. Antibodies recognizing dyskerin and GAPDH were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). GAR1 and NHP2 antibodies were obtained from Proteintech (Chicago, IL). NOP10, fibrillarin, and LC3B antibodies were from Abcam (Cambridge, MA). All other antibodies, including secondary antibodies, were obtained from Cell Signaling Technology (Danvers, MA).

Lin P., Mobasher M.E., Hakakian Y., Kakarla V., Naseem A.F., Ziai H, & Alawi F. (2015). Differential requirements for H/ACA ribonucleoprotein components in cell proliferation and response to DNA damage. Histochemistry and cell biology, 144(6), 543-558.

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Neurochemical Modulation of Neuronal Excitability

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The following chemicals were products of R&D Systems: NT, TTX, kynurenic acid, picrotoxin, SR 48692, GDP-β-S, U73122, U73343, heparin, thapsigargin, chelerythrine, bisindolylmaleimide II (Bis II), ML 133, and tertiapin-Q (TQ). Dioctanoyl phosphatidylinositol 4,5-bisphosphate (dic8-PIP₂) was purchased from Echelon Biosciences. PD149163 was product of Millipore Sigma. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals requiring dimethyl sulfoxide (DMSO) as a solvent, the concentration of DMSO was less than 0.1%. This concentration of DMSO either in the recording pipettes or in the bath had no significant effects on NT-elicited facilitation of AP firing (data not shown).

Lei S, & Hu B. (2021). Ionic and Signaling Mechanisms Involved in Neurotensin-mediated Excitation of Central Amygdala Neurons. Neuropharmacology, 196, 108714.

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Thapsigargin-Induced Calcium Signaling

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CAI, YM58493, thapsigargin were provided by Tocris (R&D Systems). ABT-737 was supplied by Selleckem and Bortezomib (Velcade®) was supplied by Millennium Pharmaceuticals. Fura-2AM was purchased from Thermo Fisher Scientific. These compounds were commonly stored as stock solutions in DMSO at −20°C.

Bonnefond M.L., Florent R., Lenoir S., Lambert B., Abeilard E., Giffard F., Louis M.H., Elie N., Briand M., Vivien D., Poulain L., Gauduchon P, & N'Diaye M. (2018). Inhibition of store-operated channels by carboxyamidotriazole sensitizes ovarian carcinoma cells to anti-BclxL strategies through Mcl-1 down-regulation. Oncotarget, 9(74), 33896-33911.

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Pharmacological Reagents for Neuronal Research

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The following chemicals were products of R&D Systems: AVP, TTX, kynurenic acid, picrotoxin, 6,7-dinitroquinoxaline-2,3-dione (DNQX), SR49059, GDP-β-S, U73122, U73343, heparin, thapsigargin, chelerythrine, bisindolylmaleimide II (Bis II), ML 133, ML 297, tertiapin-Q and tertiapin-LQ. Dioctanoyl phosphatidylinositol 4,5-bisphosphate (dic8-PIP₂) was purchased from Echelon Biosciences. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals requiring dimethyl sulfoxide (DMSO) as a solvent, the concentration of DMSO was less than 0.1%. This concentration of DMSO either in the recording pipettes or in the bath had no significant effects on neuronal activity.

Hu B., Boyle C.A, & Lei S. (2021). Roles of PLCβ, PIP2 and GIRK Channels in Arginine Vasopressin-elicited Excitation of CA1 Pyramidal Neurons. Journal of cellular physiology, 237(1), 660-674.

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Neuronal Activity Modulation Protocols

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The following chemicals were products of R&D Systems: [Thr⁴,Gly⁷]-oxytocin (TGOT), kynurenic acid, picrotoxin, GDP-β-S, U73122, U73343, heparin, thapsigargin, BAPTA, chelerythrine, bisindolylmaleimide II (Bis II), phorbol 12-myristate 13-acetate (PMA), tetrodotoxin (TTX), ML 133, ML 297, tertiapin-Q and tertiapin-LQ. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals which are only soluble in dimethyl sulfoxide (DMSO), the concentration of DMSO was less than 0.1%. This concentration of DMSO either in the recording pipettes or in the bath had no significant effects on neuronal activity.

Hu B., Boyle C.A, & Lei S. (2020). Oxytocin Receptors Excite Lateral Nucleus of Central Amygdala by PLCβ and PKC-dependent Depression of Inwardly Rectifying K+ Channels. The Journal of physiology, 598(16), 3501-3520.

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Pharmacological Toolbox for Neuronal Studies

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[Arg⁸]-vasopressin (AVP) was purchased from Bachem. The following chemicals were products of R&D Systems: TTX, kynurenic acid, picrotoxin, SR49059, GDP-β-S, U73122, U73343, heparin, thapsigargin, BAPTA, chelerythrine, bisindolylmaleimide II (Bis II), capsazepine, M084, edelfosine and pyrazolo[1,5-a]pyrimidin-7(4H)-on (QO-58). Dioctanoyl phosphatidylinositol 4,5-bisphosphate (diC8-PIP₂) was purchased from Echelon Biosciences. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals requiring dimethyl sulfoxide (DMSO) as a solvent, the concentration of DMSO was less than 0.1%. This concentration of DMSO either in the recording pipettes or in the bath had no significant effects on neuronal activity.

Boyle C.A., Hu B., Quaintance K.L, & Lei S. (2021). Involvement of TRPC5 Channels, Inwardly Rectifying K+ channels, PLCβ and PIP2 in Vasopressin-mediated Excitation of Medial Central Amygdala Neurons. The Journal of physiology, 599(12), 3101-3119.

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Detailed Chemical Source Protocol

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The following chemicals were products of R&D Systems: senktide, TTX, picrotoxin, SB218795, ML133, glibenclamide, tertiapin-Q, ML297, M084, U73122, heparin, thapsigargin, chelerythrine and bisindolylmaleimide II (Bis II). Dioctanoyl phosphatidylinositol 4,5-bisphosphate (dic8-PIP₂) was purchased from Echelon Biosciences. The other chemicals were the products of Sigma Aldrich. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals requiring dimethyl sulfoxide (DMSO) as a solvent, the concentration of DMSO was less than 0.1%.

Boyle C.A., Hu B., Quaintance K.L., Mastrud M.R, & Lei S. (2022). Ionic signaling mechanisms involved in neurokinin-3 receptor-mediated augmentation of fear-potentiated startle response in the basolateral amygdala. The Journal of physiology, 600(19), 4325-4345.

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Oxidative Stress and DNA Provocation Protocols

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For oxidative stress, hydrogen peroxide (H₂O₂) solution (30%) (Fujifilm Wako, 084‐07441) was diluted 1000‐fold with the culture medium and added to both the apex and bottom of the ALI. Cells were preincubated for 1 h with Mito‐TEMPO (Hydorate, Cayman Chemical 16621) or 2 h with the STING inhibitor, H‐151 (InvivoGen inh‐h151) and then stimulated with H₂O₂.
For intracellular DNA provocation, NCI‐H292 cells were transfected with 0.5 μg of poly(dA:dT) naked (Invivogen, tlrl‐patn) using the FuGENE HD Transfection Reagent (Promega, E2311) and incubated for 24 h. 2′3′ cGAMP (InvivoGen, tlrl‐cga23‐s) was dissolved in double‐distilled water at 1 mg/mL and stored at −20°C until use. Cells were stimulated with 2′3′ cGAMP for 16 h. MitoTEMPO (Cayman Chemical, 16621) was preadministered to cells 1 h before H₂O₂ stimulation. Thapsigargin (Cayman Chemical, 10522) was dissolved in dimethyl sulfoxide at 1 mM and stored at −20°C until use. Thapsigargin, which can trigger mitochondrial DNA release into the cytosol,¹³ was diluted with culture medium to appropriate concentrations, incubated for 4 h, and further cultured in serum‐free RPMI‐1640 for 20 h. Cells were incubated with a medium containing recombinant human transforming growth factor‐alpha (TGF‐α; R&D 239‐A‐100), an EGF receptor (EGFR) agonist for 6 h.

Nishida Y., Yagi H., Ota M., Tanaka A., Sato K., Inoue T., Yamada S., Arakawa N., Ishige T., Kobayashi Y., Arakawa H, & Takizawa T. (2023). Oxidative stress induces MUC5AC expression through mitochondrial damage‐dependent STING signaling in human bronchial epithelial cells. FASEB BioAdvances, 5(4), 171-181.

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Pharmacological Agents in Neuronal Signaling

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The following chemicals were products of R&D Systems: AVP, tetrodotoxin (TTX), kynurenic acid, picrotoxin, SR49059, TASP0390325, tolvaptan, GDP-β-S, U73122, U73343, heparin, thapsigargin, chelerythrine, bisindolylmaleimide II (Bis II), KB-R7943, capsazepine, AMG9810, AMG1629, capsaicin, ML 133, ML 297 and SCH23390. Dioctanoyl phosphatidylinositol 4,5-bisphosphate (dic8-PIP₂) was purchased from Echelon Biosciences. Drugs were initially prepared in stock solution, aliquoted and stored at −20°C. For those chemicals requiring dimethyl sulfoxide (DMSO) as a solvent, the concentration of DMSO was less than 0.1%. This concentration of DMSO either in the recording pipettes or in the bath had no significant effects on neuronal activity.

Lei S., Hu B, & Rezagholizadeh N. (2021). Activation of V1a Vasopressin Receptors Excite Subicular Pyramidal Neurons by Activating TRPV1 and Depressing GIRK Channels. Neuropharmacology, 190, 108565.

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Transfection and BET Modulation in hDPs

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hDPs were transfected with pSV3neo (ATCC no. 37150), a plasmid containing coding sequences of SV40 T-Ag and neomycin (G418)-resistance gene, as described previously [12 (link), 13 (link)]. Transfection was performed using SuperFect Transfection Reagent (Qiagen, Germany) in those cells (8 × 10⁴ cells/well) in the 24-well plates. Cell selection was performed by maintaining the transfected cells in a growth medium supplemented with 100 µg/mL G418 (Thermo Fisher Scientific, USA).
In some experiments, cells were treated with 50 mM BET (Cat. no. 107437, Sigma-Aldrich, USA). To investigate the influence of BET on calcium influx, cells were pretreated with 10 µM thapsigargin (Cat. no. 1138/1, R&D Systems, USA) or 50 µM TMB-8 (Cat. no. 53464725, Sigma-Aldrich, USA) before being stimulated with 50 mM BET (n = 6).

Kornsuthisopon C., Nantanapiboon D., Rochanavibhata S., Nowwarote N., Namangkalakul W, & Osathanon T. (2022). Betaine promotes osteogenic differentiation in immortalized human dental pulp-derived cells. BDJ Open, 8, 31.

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