Primary antibodies were: mouse monoclonal for DARPP-32 5a and 6 (ref. 58 (link); 1/5,000), calmodulin (Upstate Cell Signaling # 05–173); rabbit polyclonal for pThr34, pThr75 and pSer97 (#12438, #2301, #3401; 1:2,000; Cell Signaling); total β-adducin (#AP20545PU-N, 1:500; Santa Cruz, Acris); pSer713 of β-adducin (also reacts with pSer662 of γ-adducin and pSer724 of α-adducin; #05-587; 1:1,000; Chemicon/Millipore); β-actin (#A5316; 1:1,000; Sigma); myc-tag (#05-724; 1:500; Millipore); GFP (Abcam #ab6556, 1/1,000); pThr79-pSer81-calmodulin (Abcam, #ab194526); and pSer10 histone H3 (#06-570; rabbit; Millipore). Secondary antibodies comprised of IRDye800-conjugated anti-mouse and anti-rabbit (#610-132-121, #611-132-122; both 1:4,000; Rockland) antibodies for immunoblotting and anti-mouse CY3-conjugated antibody (#A10521; 1:400; Molecular Probes) for immunochemistry. COS-7 cells were treated with one or the combination of the following compounds, as indicated, or with dimethyl sulfoxide vehicle: Sp 5,6-DCl-cBIMPS (10 μM; BIOLOG Life Science); forskolin (100 μM; Sigma); tetradecanoylphorbol-acetate (100 nM; Sigma); okadaic acid (200 nM; Sigma); and tautomycetin (10 nM; Tocris). Mice were injected intraperitoneally with the following drugs or with their vehicle (9 g l−1 NaCl) caffeine (7.5 mg kg−1, Sigma) and cocaine (10 or 20 mg kg−1, Coper).
Tetradecanoyl phorbol acetate
Manufactured by Merck Group
Sourced in United Kingdom, Israel
Tetradecanoyl Phorbol Acetate (TPA) is a chemical compound used as a laboratory reagent. It is a phorbol ester that functions as a potent activator of protein kinase C (PKC), a family of enzymes involved in various cellular processes. TPA is commonly used in scientific research to study cell signaling pathways, cell differentiation, and other cellular mechanisms.
Automatically generated - may contain errors
Lab products found in correlation
Fetal bovine serum (fbs), by Thermo Fisher Scientific (3 mentions)
4 6 diamino 2 phenylindole dapi, by Merck Group (2 mentions)
Fetal calf serum (fcs), by Merck Group (2 mentions)
Puromycin, by Avantor (2 mentions)
Leupeptin, by Avantor (2 mentions)
Phenyl methyl sulfonyl fluoride (pmsf), by Avantor (2 mentions)
Aprotinin, by Avantor (2 mentions)
Pepstatin a, by Avantor (2 mentions)
Doxycycline dox, by Sangon (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Cholera toxin, by Merck Group (2 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (2 mentions)
Hepes, by Thermo Fisher Scientific (2 mentions)
2 mercaptoethanol, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640 medium, by Merck Group (2 mentions)
Anti mouse cy3 conjugated antibody, by Thermo Fisher Scientific (1 mentions)
Forskolin, by Merck Group (1 mentions)
Myc tag, by Merck Group (1 mentions)
Okadaic acid, by Merck Group (1 mentions)
Ap20545pu n, by Santa Cruz Biotechnology (1 mentions)
16 protocols using tetradecanoyl phorbol acetate
1
Phosphorylation and Regulation of Neuronal Proteins
2
Culturing Hepatocellular Carcinoma Cell Lines
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The cultured conditions for HCC340 and HepG2 cells were the same as described in our previous reports24 (link),35 (link). HCC 340 is a patient-derived hepatocellular carcinoma cell line from Buddhist Tzu Chi Hospital, Taiwan24 (link). Tetradecanoyl phorbol acetate (TPA) was purchased from Sigma-Aldrich (Poole, UK). The snail expression plasmid, p-Snail, driven by CMV promoter within the pcDNA3 vector, is a gift from Dr. Cheng K.K. in Tzu Chi university.
3
Molecular Signaling Pathway Analysis
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Tetradecanoyl phorbol acetate (TPA), poly-L-Lysine (PLL) and 4,6-diamino-2-phenylindole (DAPI) were purchased from Sigma-Aldrich (Rehovot, Israel). Albumin bovine serum (BSA) was purchased from MP biomedical (OH, USA). The BRAF inhibitor PLX4032 and MEK inhibitor Trametinib (GSK1120212) were purchased from SelleckChem (Huston, TX). CellTiter-Glo reagent was purchased from Promega (Madison, WI).
4
Intracellular Cytokine Profiling by Flow Cytometry
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Intracellular cytokines were detected by flow cytometry. Briefly, T cells (1×106/mL) were stimulated with immobilized anti‐CD3 (1 μg/mL; OKT3, eBioscience) and tetradecanoylphorbol acetate (10 ng/mL; Sigma) for 6 hours in complete medium. Prior to the culture, the plates were centrifuged for 5 minutes at 800g. Three hours after activation, monensin (10 μg/mL; Sigma) was added. T cells were then collected, washed in phosphate‐buffered saline, and fixed with 2% formaldehyde. After fixation, T cells were permeabilized in phosphate‐buffered saline supplemented with 2% Fetal Calf serum and 0.5% saponin (Sigma). Permeabilized T cells were incubated with anti‐IL‐17, anti‐IL‐4, anti‐interferon‐γ, anti‐IL‐10, or anti‐FoxP3 monoclonal antibodies. All monoclonal antibodies were obtained from BD PharMingen. After washing, cells were analyzed using LSRII Fortessa™ cell analyzer (BD Biosciences, CA), and data were analyzed with Flowjo software. Quadrant markers were set accordingly to isotype‐matched controls.
5
Resveratrol Analogues in Melanocyte Culture
6
Molecular Interaction Detection Toolkit
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Tetradecanoyl phorbol acetate (TPA), anisomicin, Polyethylenimine (PEI) and 4'6-diamino-2phenylindole (DAPI) were obtained from Sigma (Rehovot, Israel). A/G beads were purchased from Santa Cruz Biotechnology (Dallas, TX, USA). Albumin bovine serum (BSA) was purchased from MP niomedical (Solon, OH, USA). Si RNAs and Dharmafect were from Thermo Fisher Scientific (CO, USA). Proximity ligation assay kit was from Olink Bioscience (Uppsala, Sweden). NBT/BCIP developing substrates were purchased from Promega (WI, USA), Calf Intestinal Phosphatase (CIP) was from New England Biolabs (MA, USA).
7
Immunoprecipitation and Immunoblotting Assay
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Antibodies to EBNA1 (sc-57719, Santa cruz), FLAG (M2, Sigma), SUMO1 (Y299, Abcam), SUMO2/3 (for IB/IF, EPR4602, Abcam; for IP, fsc-393144, Santa cruz), STUB1 (EPR4447, Abcam), KAP1 (20C1, Abcam), GST (12G8, M20007, Abmart) and GAPDH (G8140-01, US Biological) were used according to the manufacturers specifications. The monoclonal antibody anti-myc (9E10) and HA (12CA5) were prepared from hybridoma cultures stored in the laboratory. Tetradecanoyl Phorbol Acetate (TPA) was purchased from Sigma and sodium butyrate from J&K Corporation. Proteasome inhibitors PMSF, Leupeptin, Aprotinin, Pepstatin A and Puromycin were purchased from Amresco.
8
Snail Signaling Pathway Activation
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Tetradecanoyl phorbol acetate (TPA) was purchased from Sigma-Aldrich (Poole, UK). The monoclonal Snail (C15D3) antibody; and the antibodies against EGR1, SP1, MMP9, ZEB1, LEF1, Fibronectin, COX2, COL1A1, Histone H3, and GAPDH were purchased from Cell Signaling (Beverly, MA, USA) and Santa Cruz Biotechnology (Santa Cruz, CA, USA), respectively. The snail expression plasmid, p-Snail, driven by CMV promoter within the p-cDNA3 vector, was a gift from Dr. Cheng K.K., Tzu Chi University.
9
Cell Culture and Serum Starvation Protocol
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Human U-2OS osteosarcoma cells [70 (link)] expressing the murine ecotropic receptor were kindly provided by D. Shvarts (Utrecht Medical Center, Utrecht, The Netherlands), HeLa cervical carcinoma cells [71 (link)] by M. Koritzinsky (MAASTRO, Maastricht, The Netherlands). Normal human fibroblasts BJ [72 (link)] and TIG3 [73 (link)] were obtained through collaboration; normal human TIG3 fibroblasts expressing the murine ecotropic receptor: courtesy D. Peeper (Netherlands Cancer Institute, Amsterdam, The Netherlands). Cells were cultured under standard conditions in medium supplemented with 10% fetal calf serum and antibiotics (100 units/ml penicillin and 100 μg/ml streptomycin; Gibco). Serum starvation: 0.05% FCS (cancer cell lines) and 0.1% (human fibroblasts) for 48 hrs. Mitogenic stimulation: supplementation of 15% FCS and 100ng/ml tetradecanoyl phorbol acetate (TPA; Sigma-Aldrich, St. Louis, MO, USA) for the indicated duration.
10
Antibody Detection of KSHV Proteins
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A mouse monoclonal (PG-M3) and rabbit polyclonal antibodies (Ab; H-238) against PML were purchased from Santa Cruz Biotechnology and used to detect endogenous and exogenous PML. Rabbit polyclonal anti-hDAXX (HPA008736) and anti-SP100 (HPA016707) Abs were purchased from Sigma-Aldrich. Mouse monoclonal antibodies to RTA, K8 (K-bZIP), K5, K3, K9 (vIRF1), ORF59, and K8.1 were developed previously in our laboratory and rabbit polyclonal Ab against gB was a kind gift from Bala Chandran (Akula et al., 2001 (link); Okuno et al., 2002 (link)). A rat monoclonal anti-LANA (HHV-8 ORF73) Ab was purchased from Advanced Biotechnologies (Cat# 13-210-100). An anti-Halo Tag® mouse monoclonal Ab (Cat# G9211, Promega, United States) was used to detect exogenous Halo-tagged PML. A mouse monoclonal antibody (Sigma Cat# T5201) was used to detect β-tubulin. Horseradish peroxidase (HRP)-labeled goat polyclonal anti-rabbit, anti-mouse, and anti-rat immunoglobulins (Dako, Denmark) were used as secondary antibodies in Western blot analysis. Anti-mouse, anti-rat, and anti-rabbit IgG conjugated Alexa Fluor® 488 and/or Alexa Fluor® 546 (Molecular Probes, United States) were used as secondary antibodies for immunofluorescence assays (IFA). Tetradecanoyl phorbol acetate (TPA; Sigma, Japan) and sodium butyrate (NaB; Sigma, Japan) were used to induce lytic replication of KSHV.
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