Antibodies against HA-Tag Rabbit (C29F4, #3724), HA-Tag mouse (6E2, #2367), acetyl-lysine (#9441), p-P38 (Thr180/Tyr182) (D3F9, #4511), P38(#9212), p-ERK (E-4, #sc-7383), ERK (#4696), p-AKT (Ser473, 193H12, #4058), AKT (#9272), p-mTOR (Ser2448, #2971), mTOR (#2983), p-p70 S6K (Thr421/Ser424, #9204), S6K (#9202), and UBC9 (D26F2, #4786) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against SENP1 (A1260) were purchased from ABclonal (Woburn, MA, USA). Antibodies against Flag-tag (F1804), MG132, and cycloheximide were purchased from Sigma-Aldrich (St. Louis, MO, USA). Antibodies against Sirt2 (#ab67299) and SUMO1 (#Y299) were purchased from Abcam (Cambridge, UK). Puromycin (P8230) and rapamycin (R8140) were purchased from Solarbio (Beijing, China). Regents of SB202190 (#S1077), LY294002 (#S1105), NAD+ (#S2518), trichostatin A (TSA) (#S1045), nicotinamide (NAM) (#S1899), and protease inhibitor cocktail (EDTA-Free,100 × in DMSO) were obtained from Selleck (Houston, TX, USA). The KOD-plus-mutagenesis kit was purchased from Toyobo (Osaka, Japan). Protein A/G magnetic beads were purchased from Thermo Fisher Scientific (Waltham, MA, USA). Ni-NTA beads were purchased from Qiagen (Hilden, Germany).
Trichostatin a tsa
Manufactured by Selleck Chemicals
Sourced in United States
Trichostatin A (TSA) is a histone deacetylase (HDAC) inhibitor. It is a laboratory reagent used in the study of epigenetic mechanisms and cellular processes.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
L1200 kinase inhibitor library, by Selleck Chemicals (2 mentions)
Mk 2461, by Selleck Chemicals (2 mentions)
Click it edu cell proliferation kit for imaging with alexa fluor 647 dye, by Thermo Fisher Scientific (2 mentions)
Hct116, by American Type Culture Collection (2 mentions)
Sw480, by American Type Culture Collection (2 mentions)
Sw620, by American Type Culture Collection (2 mentions)
Fhs 74 int, by American Type Culture Collection (2 mentions)
U0126, by Selleck Chemicals (2 mentions)
P mtor ser2448, by Cell Signaling Technology (1 mentions)
Protein a g magnetic bead, by Thermo Fisher Scientific (1 mentions)
Ni nta bead, by Qiagen (1 mentions)
Kod plus mutagenesis kit, by Toyobo (1 mentions)
Cycloheximide (chx), by Merck Group (1 mentions)
Protease inhibitor cocktail, by Selleck Chemicals (1 mentions)
Rapamycin, by Solarbio (1 mentions)
Puromycin p8230, by Solarbio (1 mentions)
Acetyl lysine, by Cell Signaling Technology (1 mentions)
Ly294002, by Selleck Chemicals (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
15 protocols using trichostatin a tsa
1
Comprehensive Antibody Reagents for Cellular Signaling
2
Kinase Inhibitor Screen in Zebrafish
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In our initial small molecule screen, dechorionated olig2:egfp larvae were treated with 10 μm kinase inhibitor in 1% DMSO in PTU egg water from 24 to 76 h post-fertilization (hpf). Kinase inhibitors used were 1 of 430 kinase inhibitors from the L1200 Kinase Inhibitor Library (Selleck Chem), MK-2461 (Selleck Chem), or Trichostatin-A (TSA) (Selleck Chem). Control siblings were treated with 1% DMSO in PTU egg water. The small molecule screen was conducted in triplicate.
For the EdU incorporation assay, larvae were treated with 0.4 mM EdU in 4% DMSO from 70 to 74 hpf in PTU egg water at 28.5 °C then fixed for 1 h in 4% PFA at RT. Larvae were washed for 5 min with 1X PBSTX, 5 min in DWTX, then permeabilized with cold acetone for 10 min at −20°C and stained for EdU using the Click-it EdU Cell Proliferation kit for Imaging with Alexa Fluor 647 dye (ThermoFisher), as detailed in the kit protocol. Click-it reaction was performed for 1 h at RT and thoroughly washed overnight with PBSTX prior to imaging.
For the EdU incorporation assay, larvae were treated with 0.4 mM EdU in 4% DMSO from 70 to 74 hpf in PTU egg water at 28.5 °C then fixed for 1 h in 4% PFA at RT. Larvae were washed for 5 min with 1X PBSTX, 5 min in DWTX, then permeabilized with cold acetone for 10 min at −20°C and stained for EdU using the Click-it EdU Cell Proliferation kit for Imaging with Alexa Fluor 647 dye (ThermoFisher), as detailed in the kit protocol. Click-it reaction was performed for 1 h at RT and thoroughly washed overnight with PBSTX prior to imaging.
3
Graphene Oxide Nanoparticle Imaging Protocol
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Graphene oxide nano-colloids (GON) dispersed in H2O (2 mg/mL), paraformaldehyde (PFA), methanol and all the reagents used for solutions were purchased from Sigma Aldrich. Bovine Serum Albumin (BSA) and goat serum were purchased from Santa Cruz Biotechnology. The signaling pathway inhibitors Dabrafenib/GSK2118436 (DAB) and Trichostatin A (TSA) were purchased from Selleckchem (www.Selleckchem.com ).
4
Breast Cancer Cell Line Culture and Inhibitor Treatment
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MCF-7 and MDA-MB-231 human breast cancer cell lines were purchased from the American Type Culture Collection and were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% fetal bovine serum (FBS; Thermo-Fischer Scientific, Waltham, MA, USA) and a 1% antibiotic–antimycotic solution (Thermo-Fischer Scientific, Waltham, MA, USA) in a humidified 5% CO2 atmosphere at 37 °C. Cells were treated with TNF-α (Sigma-Aldrich, St. Louis, MD, USA) as the indicated conditions. The Lipofectamine 2000 transfection reagent was purchased from Thermo-Fischer Scientific. Z-VAD-FMK, a pan-caspase inhibitor and Z-DQMD-FMK, a caspase-3 inhibitor were obtained from R&D systems (Minneapolis, MN, USA). MS-275, an HDAC1 & 3 inhibitor and trichostatin A (TSA), a pan-HDAC inhibitor were purchased from Selleckchem (Houston, TX, USA).
5
Fluorimetric HDAC2 Activity Assay
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HDAC2 activity was determined using a fluorimetric assay at BPS Bioscience (San Diego, CA, USA). Compounds were dissolved in DMSO (Sigma), and serial dilutions were further diluted in HDAC assay buffer (BPS). These working solutions were incubated in duplicate at room temperature for 3 h in a mixture containing HDAC assay buffer, BSA (Fisher Scientific), and recombinant HDAC2 (BPS). The enzymatic reactions were initiated by the addition of a fluorogenic, acetylated peptide substrate (BPS) and proceeded for 30 min at 37 °C. Then, HDAC assay developer (BPS) was added, and after further incubation at room temperature, fluorescence intensity was measured at an excitation of 360 nm and an emission of 460 nm using a Tecan Infinite M1000 microplate reader. Trichostatin A (TSA, Selleckchem, Houston, TX, USA) was used as a reference inhibitor.
6
Pharmacological Inhibition of Kinases and Histone Deacetylases
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H-89 dihydrochloride (PKA/PKB/AKT1 kinase inhibitor), 3-aminobenzoic acid ethyl ester (tricaine), and rifampicin were purchased from Sigma-Aldrich, Zwijndrecht, The Netherlands. H-89 analog 97i was synthesized by the Leiden Academic Center for Drug Research, Division of Medicinal Chemistry, Leiden University, Leiden, The Netherlands. Pan-HDAC inhibitor Trichostatin A (TSA) and class IIa HDAC inhibitors TMP195 and TMP269 were purchased from Selleckchem, Munich, Germany. Hygromycin B was acquired from Life Technologies-Invitrogen, Bleiswijk, The Netherlands. Recombinant human IFN-γ protein was acquired from R&D Systems, Wiesbaden, Germany.
7
Treating Viral Myocarditis in Mice
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BALB/c mice in each group were infected by an intraperitoneal injection with 1 × 103 TCID50 CVB3 on day 0. To examine the therapeutic effects of suberoylanilide hydroxamic acid (SAHA; Selleck), the solution of SAHA in 2-hydroxypropyl-β-cyclodextrin (HOP-β-CD; Sigma-Aldrich) was prepared as previously described (22 (link)). Mice were orally administered daily with 50 mg/kg of body weight of SAHA, starting from the day of virus inoculation. In addition, trichostatin A (TSA; Selleck) was dissolved in dimethyl sulfoxide (DMSO) and injected intraperitoneally into mice at 0.5 mg/kg/day. Ribavirin (Selleck) was dissolved in phosphate-buffered saline (PBS) and injected intraperitoneally into mice at 100 mg/kg/day.
8
Epigenetic Regulation in Ovarian Cancer
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The activity of DNMT was determined by EpiQuik DNMT Activity/Inhibition Assay Ultra Kit (Colorimetric) (Epigentec Group Inc., Farmingdale, NY, USA) in ovarian cancer cells treated with ginsenoside Rg3 (0, 50, 100 and 200 µg/ml) or 50 µg/ml 5-aza-dc (as a negative control). The samples were first co-incubated with cytimidine substrate and then with 5′-methyl-cytimidine antibody. The activity of HDAC was determined using Epigenase HDAC Activity/Inhibition Direct Assay Kit (Colorimetric) (Epigentec Group Inc.) in ovarian cancer cells treated with ginsenoside Rg3 (0, 50, 100 and 200 µg/ml) or 500 ng/ml HDAC inhibitor trichostatin A (TSA) (Selleck Chemicals, Houston, TX, USA) (as a negative control). The samples were co-incubated with acetylated substrate and then with photographic developer. The OD values at 450 nm were measured using a MultiSkan 1500 microplate reader (Thermo Fisher Scientific, Inc.).
9
Establishing Highly Invasive CRC Cell Line
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Human colon cancer cell lines SW480, SW620, HCT116, RKO, and normal small intestine epithelial cell FHs 74Int were purchased from ATCC (Rockville, MD), and the CRC cells were maintained in 1640 medium supplemented with 10% fetal bovine serum (FBS, Gibco, CA, USA) at 37°C with 5% CO2. HCT116 highly invasive cell line (HCT116-i8) was established using repeated invasion assays as previously described 21 (link). All the cell lines were tested for mycoplasma contamination. U0126 and trichostatin A (TSA) were purchased from Selleck Chemicals (cat#S1102 and cat#S1045, Houston, TX, USA). U0126 (10 mM, 10 mg in 2.35 mL dimethyl sulfoxide (DMSO)), and TSA (10 mM, 5 mg in 1.65 mL DMSO) were respectively dissolved for in vitro usage.
10
Colorectal Cancer Cell Line Maintenance
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The human colorectal cancer cell lines RKO and DLD1 (ATCC, LGC Standards, Middlesex, UK) and their CPT-11 resistant derivative single clones were maintained in a 5% CO2 incubator at 37°C and grown in Gibco RPMI media (Life Technologies, Paisley, UK) supplemented with 10% FCS and 2 mM l -glutamine. Irinotecan hydrochloride (CPT-11), 5-fluorouracil (5-FU), oxaliplatin, hydroxyurea (HU), camptothecin (CPT), doxorubicin, etoposide and SN-38 were obtained from Sigma-Aldrich (Gillingham, UK) whilst the PARP inhibitor, olaparib (AZD2281) was obtained from Stratech Scientific Limited (Suffolk, UK). Trichostatin A (TSA) and Panobinostat (LBH-589) were obtained from Selleckchem.
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