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Tetradecanoylphorbol Acetate

Tetradecanoylphorbol acetate (TPA) is a potent tumor-promoting agent and a widely used activator of protein kinase C.
It is frequently utilized in cell biology and cancer research to investigate signaling pathways, cell differentiation, and tumor progression.
PubCompare.ai can optimize your TPA research by leveraging AI to identify the best protocols from scientific literature, preprints, and patents, enhancing reproducibility and accuracy.
Discover how this tool can help you find the optimal products and procedures for your TPA experiments and take your research to the next level.

Most cited protocols related to «Tetradecanoylphorbol Acetate»

Compounds were purchased from Chemdiv, San Diego CA: CK-0944636 (Catalog number 8012-5103), CK-0993548 (Catalog number K205-1650), CK-0944666 (Catalog number 8012-5153) and CK-0157869 (Catalog number K205-0942). We purified native Arp2/3 complex from human platelets12 (link), bovine thymus6 (link), Schizosaccharomyces pombe13 (link) and Saccharomyces cerevisiae (Supplemental methods), actin from chicken skeletal muscle14 (link) and recombinant HsWASp, WASp105-502, WASp-VCA and Cdc4212 (link), N-WASp-VCA 428-505 (Supplemental methods), GST-ActA 36-170 (Supplemental methods) and S. pombe Cdc12p(FH2)-His 973-139015 (link) from E. coli. We used standard assays to measure polymerization of pyrenyl-actin16 (link) and to visualize actin filaments by fluorescence microscopy17 (link). Binding of etheno-ATP to Arp2/3 complex was performed as described previously with slight modifications18 (link). We crystallized BtArp2/3 complex7 (link) with either 0.5 mM CK-548 or 1 mM CK-636 in DMSO or soaked these compounds into crystals for 24 hours before freezing in liquid nitrogen. Diffraction data were collected at beamline X29A at Brookhaven National Laboratories. SKOV3 cells were infected with Listeria monocytogenes and fixed with 2% formaldehyde, permeabilized with 0.1% Triton-X in PBS, stained with Listeria antibody (US Biologics, Cleveland, Ohio) and Alexa Fluor 568 phalloidin (Molecular Probes, Eugene, OR), and imaged by fluorescence microscopy. We used an Isodata threshold on background-subtracted images of Listeria to isolate individual bacterium and measure the ratio of colocalized actin to Listeria fluorescence. Monocyte THP-1 cells were differentiated in 50 nM phorbol myristate acetate (Sigma-Aldrich-Fluka) to form podosomes before treatment with compounds. Black molly keratocytes19 (link) were observed by time-lapse phase contrast microscopy.
Publication 2009
Actin-Related Protein 2-3 Complex Actins alexa 568 Bacteria Biological Assay Biological Factors Cattle Cells Chickens CK-0944636 CK-0944666 CK-0993548 Escherichia coli Fluorescence Formaldehyde Homo sapiens Immunoglobulins Isoenzyme CPK MB Listeria Listeria monocytogenes Microfilaments Microscopy, Fluorescence Microscopy, Phase-Contrast Molecular Probes Molly Monocytes Nitrogen Phalloidine Podosomes Polymerization Saccharomyces cerevisiae Schizosaccharomyces Schizosaccharomyces pombe Skeleton Sulfoxide, Dimethyl Tetradecanoylphorbol Acetate THP-1 Cells WASL protein, human WAS protein, human
Compounds were purchased from Chemdiv, San Diego CA: CK-0944636 (Catalog number 8012-5103), CK-0993548 (Catalog number K205-1650), CK-0944666 (Catalog number 8012-5153) and CK-0157869 (Catalog number K205-0942). We purified native Arp2/3 complex from human platelets12 (link), bovine thymus6 (link), Schizosaccharomyces pombe13 (link) and Saccharomyces cerevisiae (Supplemental methods), actin from chicken skeletal muscle14 (link) and recombinant HsWASp, WASp105-502, WASp-VCA and Cdc4212 (link), N-WASp-VCA 428-505 (Supplemental methods), GST-ActA 36-170 (Supplemental methods) and S. pombe Cdc12p(FH2)-His 973-139015 (link) from E. coli. We used standard assays to measure polymerization of pyrenyl-actin16 (link) and to visualize actin filaments by fluorescence microscopy17 (link). Binding of etheno-ATP to Arp2/3 complex was performed as described previously with slight modifications18 (link). We crystallized BtArp2/3 complex7 (link) with either 0.5 mM CK-548 or 1 mM CK-636 in DMSO or soaked these compounds into crystals for 24 hours before freezing in liquid nitrogen. Diffraction data were collected at beamline X29A at Brookhaven National Laboratories. SKOV3 cells were infected with Listeria monocytogenes and fixed with 2% formaldehyde, permeabilized with 0.1% Triton-X in PBS, stained with Listeria antibody (US Biologics, Cleveland, Ohio) and Alexa Fluor 568 phalloidin (Molecular Probes, Eugene, OR), and imaged by fluorescence microscopy. We used an Isodata threshold on background-subtracted images of Listeria to isolate individual bacterium and measure the ratio of colocalized actin to Listeria fluorescence. Monocyte THP-1 cells were differentiated in 50 nM phorbol myristate acetate (Sigma-Aldrich-Fluka) to form podosomes before treatment with compounds. Black molly keratocytes19 (link) were observed by time-lapse phase contrast microscopy.
Publication 2009
Actin-Related Protein 2-3 Complex Actins alexa 568 Bacteria Biological Assay Biological Factors Cattle Cells Chickens CK-0944636 CK-0944666 CK-0993548 Escherichia coli Fluorescence Formaldehyde Homo sapiens Immunoglobulins Isoenzyme CPK MB Listeria Listeria monocytogenes Microfilaments Microscopy, Fluorescence Microscopy, Phase-Contrast Molecular Probes Molly Monocytes Nitrogen Phalloidine Podosomes Polymerization Saccharomyces cerevisiae Schizosaccharomyces Schizosaccharomyces pombe Skeleton Sulfoxide, Dimethyl Tetradecanoylphorbol Acetate THP-1 Cells WASL protein, human WAS protein, human
The melanoma cells listed in Table 1 were isolated from primary and metastatic lesions and the normal melanocytes from a giant nevus, all excised to improve patient quality of life. The specimens were collected with participants’ informed consent according to Health Insurance Portability and Accountability Act (HIPAA) regulations with Human Investigative Committee protocol. The melanoma cells from advanced lesions were grown in OptiMEM (Invitrogen, Carlsbad, CA, USA) supplemented with antibiotics and 5% fetal calf serum (basal medium). WW165 cells were cultured in basal medium supplemented with 3-isobutyl-1-methylxanthine (IBMX; Sigma-Aldrich, St Louis, MO, USA), and YULOVY, YUFULO and YUREEL-NV with recombinant fibroblast growth factor 2 (bFGF; Promega, Madison, WI, USA), IBMX and 12-O-tetradecanoylphorbol-13-acetate (TPA; Sigma), ingredients required for optimal proliferation (Böhm et al., 1995 (link); Hoek et al., 2004 (link)). YUSIT1, 501 mel, YUGEN8 and WW165 were long-term cultures and the rest were early, short-term passage (passage 2–15). Keratinocytes were cultured from newborn foreskins in EpiLife medium (Cascade Biologics, Portland, OR, USA) and used during the first passage. BRAF, NRAS and PTEN mutations were identified by Sanger sequencing of gene-specific amplicons (Tables S1).
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Publication 2010
1-Methyl-3-isobutylxanthine Antibiotics, Antitubercular Biological Factors BRAF protein, human Cells Fetal Bovine Serum Fibroblast Growth Factor 2 Foreskin Genes Giant pigmented hairy nevus Homo sapiens Infant, Newborn Keratinocyte Melanoma methylxanthine Mutation NRAS protein, human Patients Promega PTEN protein, human Tetradecanoylphorbol Acetate
At the PP stage (day 12 of differentiation), cells were washed with PBS, incubated with TrypLE at 37°C for 5–6 min and carefully resuspended in DMEM/F12 resulting in clumps of 3–10 cells. After centrifugation at 400 g for 5 min, the pellet was washed in BE3 medium, centrifuged again and resuspended in precooled day 12 medium (detailed above) supplemented with 10 µM Rock inhibitor. The cell suspension was mixed on ice with growth factor reduced matrigel at a 1:3 ratio and 25 µL were transferred to a 48-well plate (NunclonΔSurface). Following incubation for 10 min at 37°C, the solidified drop of matrigel was overlayed with 200 µL of day 12 medium (see above) containing 10 µM Rock inhibitor (always added for the first 4 days in three-dimensional (3D) culture), which was replaced the next day. On day 14, differentiation was continued with 10 ng/mL FGF2 and 10 µM Rock inhibitor in BE3 medium. From day 18 on, organoids were cultured in BE3 with 10 ng/mL FGF2 and 10 mM nicotinamide (NA) (Sigma) (referred to as ‘FN’). Medium was changed every 2–3 days. Alternatively, organoids were generated by replating PPs in suspension in ultra-low-attachment plates (Corning). To prevent aggregation, cell clusters were triturated after 1 hour and again after 2 days. For changing media, half of the fluid was pipetted off and replaced with fresh differentiation media. Medium conditions were identical to the matrigel-based culture. For suspension cultures, another medium (referred to as ‘FEPC’) was tested (based on conditions published for mouse embryonic PPs26 (link)) composed of DMEM/F12, 10% knockout serum replacement (KOSR) and 0.1 mM β-mercaptoethanol supplemented with 50 ng/mL FGF10, 25 ng/mL epidermal growth factor (EGF) (novoprotein), 2 µM CHIR99021 and 16 nM phorbol myristate acetate (Sigma). For passaging, matrigel was scraped off, pipetted in order to mechanically dissociate the organoids into small clumps, collected in a 15 mL falcon, and further processed as described above. Organoids were passaged every 10–14 days and cultured at 5% CO2 and 37°C.
Publication 2016
2-Mercaptoethanol Cells Centrifugation Chir 99021 Embryo Epidermal growth factor FGF10 protein, human Fibroblast Growth Factor 2 Growth Factor matrigel Mus Niacinamide Organoids Serum Tetradecanoylphorbol Acetate
For flow cytometry analysis, 5 mice in each group were anesthetized. The lumbar spinal cord and lymph nodes were carefully dissected and dissociated at the time of the peak clinical score as previously described [23 (link), 33 (link), 34 (link)]. A single-cell suspension was prepared and fixed with 2 % paraformaldehyde for surface cell analysis. Cells were then washed with 2 % FBS in PBS, incubated with mouse anti-rat CD32 (BD Biosciences, San Jose, CA, USA) for 10 min to block the Fc receptor and washed twice with 2 % FBS in PBS. For surface cell analysis, cells were incubated with APC anti-mouse CD11b (OX-42; Biolegend, San Diego, CA, USA), PE anti-mouse CD45 (OX-1; Biolegend), APC anti-mouse CD4 (OX-35, BD Biosciences), and PE anti-mouse CD8a (OX-8, BD Biosciences) for 30 min at 4 °C. Microglia and macrophages were identified based on their relative CD45 expression levels [54 (link)]. Briefly, after acquiring unstained and single color control samples to calculate compensation matrix, we acquired 1 × 104 events within the combined gate based on physical parameters [forward scatter (FSC) and side scatter (SSC)]. CD11b+/CD45+(low) cells and CD11b+/CD45+(high) cells were gated as resident microglia and macrophages, respectively [54 (link)]. For intracellular cell analysis, cells were restimulated with phorbol-12-myristate-13-acetate and ionomycin and Golgistop in RPMI media. After 5 h, cells were stained with PerCP-Cy 5.5 anti-mouse CD4 (RM4-5; BD Biosciences), FITC anti-mouse IFN-γ (XMG1.2; BD Biosciences), PE anti-mouse IL-17A (TC11-18H10; BD Biosciences), PE anti-mouse IL-4 (11B11; BD Biosciences), PE anti-mouse CD25 (PC61.5; FJK-16 s; eBioscience), and APC anti-mouse/rat Foxp3 (FJK-16 s; eBioscience) (Additional file 2) for 30 min at 4 °C. The cells were washed twice with 2 % FBS in PBS and used for flow cytometry. To identify CD4+ T cell populations, we first gated on cells (1 × 104) based on FSC and SSC properties. CD4+ T cells were used to analyze populations of Th1, Th2, Th17, and Treg cells on CD4. Three-color staining was performed for one cell for simultaneous analysis. Intracellular cytokine results were indicated as percentages within the CD4+ population. Data were collected on a FACS Calibur flow cytometer (BD Biosciences) and analyzed using Cell Quest Pro software (BD Biosciences).
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Publication 2016
Cardiac Arrest CD4 Positive T Lymphocytes Cells Cytokine Fc Receptor Flow Cytometry Fluorescein-5-isothiocyanate IFNG protein, mouse IL2RA protein, human IL17A protein, human Ionomycin ITGAM protein, human Lumbar Cord Macrophage Microglia Mus Nodes, Lymph paraform Physical Examination Protoplasm Regulatory T-Lymphocytes Tetradecanoylphorbol Acetate Th17 Cells

Most recents protocols related to «Tetradecanoylphorbol Acetate»

Example 4

Peripheral blood mononuclear cells (PBMCs) were isolated from freshly collected whole blood from Ulcerative Colitis (UC) and Crohn's Disease (CD) patients, by conventional density gradient centrifugation. To induce CD30L expression on primary lymphocytes, the isolated cells were stimulated overnight with Phorbol 12-myristate 13-acetate (PMA) and ionomycin. The next day, the stimulated cells, along with non-stimulated cells kept as control, were collected, washed and incubated on ice with increasing concentrations of fluorescently labeled anti-CD30L antibodies or isotype control (from 0.001 nM to 60 nM). After washing to remove unbound antibodies, the cells were fixed in a paraformaldehyde solution and analyzed by flow cytometry to quantify cell surface antibody binding. Typical results from this assay are shown in Table 7.

TABLE 7
Binding to primary lymphocytes from UC and
CD patients stimulated with PMA/ionomycin.
Patient 1Patient 2Patient 3Patient 4
(CD, #04-021)(UC, #03-041)(UC, #02-180)(CD, #01-051)
AntibodyBindingBindingBindingBinding
CloneEC50 (nM)EC50 (nM)EC50 (nM)EC50 (nM)
Ref1 (HC of3.633.123.072.82
SEQ ID NO:
768 & LC of
SEQ ID NO:
769)
17.455.435.574.53
22.401.551.711.72

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Patent 2024
Anti-Antibodies Antibodies APEX1 protein, human Biological Assay BLOOD Cells Centrifugation, Density Gradient Clone Cells Crohn Disease Flow Cytometry Immunoglobulin Isotypes Immunoglobulins Ionomycin Lymphocyte paraform Patients PBMC Peripheral Blood Mononuclear Cells Tetradecanoylphorbol Acetate Ulcerative Colitis
For intracellular staining, short-term reactivation of cryopreserved splenocytes and subsequent mass cytometry analysis were performed as described previously94 (link). In short, cells were kept at −80 °C for less than 2 months before thawing in a 37 °C water bath. Cells were then immediately resuspended in cell culture medium supplemented with 1:10,000 benzonase (Sigma-Aldrich), and centrifuged at 300 g for 7 min at 24 °C. Samples were then left overnight at 37 °C before restimulation with 50 ng/mL phorbol 12-myristate 13-acetate (Sigma-Aldrich) and 500 ng/mL ionomycin (Sigma-Aldrich) in the presence of 1× brefeldin A (BD Biosciences) and 1× monensin (Biolegend) for 4 h at 37 °C. For splenocytes, one anchor sample to correct batch effect among different acquisitions and one non-stimulated control sample were also included. For both PBMCs and reactivated cryopreserved splenocytes, surface staining was performed for 30 min at 4 °C. To identify dead cells, 2.5 μM cisplatin (Sigma-Aldrich) was added for 5 min on ice. To minimize technical variability, an equal number of cells from each sample were barcoded using Cell-ID 20-Plex (Fluidigm). Cells from all samples were then combined in one single tube. The combined sample was then processed for Live/Dead and surface staining. For intracellular cytokine staining of reactivated cryopreserved splenocytes, after surface staining, cells were fixed and permeabilized with FOXP3 staining kit (Invitrogen) and stained for intracellular markers and cytokines. The antibodies used are reported in Supplementary Table 2. Because CD4 molecules are internalized during phorbol 12-myristate 13-acetate/ionomycin stimulation95 (link), the anti-CD4 antibody was used in both the surface staining and the intracellular staining steps. After washing, the combined sample was incubated with 4% PFA overnight at 4 °C. Prior to acquisition in a Helios™ II CyTOF® system, samples were washed with cell staining buffer and mass cytometry grade water. Multidimensional datasets were analyzed using FlowJo and R (R Core Team, 2017).
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Publication 2023
Antibodies Antibodies, Anti-Idiotypic Bath Benzonase Brefeldin A Buffers CD4 Antigens Cell Culture Techniques Cells Cisplatin Culture Media Cytokine Ionomycin Monensin Protoplasm Tetradecanoylphorbol Acetate
Freund’s complete adjuvant (FCA) (F5881), lipopolysaccharide (LPS) (L2880) and phorbol 12-myristate 13-acetate (PMA) (P1585) were obtained from Sigma Chemicals (Louis, MO, USA). Sinomenine (S2359) was purchased from Selleck Chemicals (Shanghai, CHN). Anti-MPO (ab9535), anti-NE (ab21595), anti-Ly6G (ab25377), anti-CitH3 (ab5103), anti-IL-6 (ab208113), anti-LC3B (ab48394), anti-Beclin-1 (ab207612), anti-GAPDH (ab181602), anti-ERK1/2 (ab115799), anti-rabbit IgG secondary (ab6721) and goat anti-rabbit IgG H&L (Alexa Fluor® 555) secondary antibodies (ab150086) were obtained from Abcam (Cambridge, MA, USA). The anti-p65 (3033S), anti-phospho-p65 (Ser536) (6956S), anti-SAPK/JNK (9252S), anti-phospho-SAPK/JNK (Thr183/Tyr185) (9251S), anti-phospho-ERK1/2 (Thr202/Tyr204) (9101S), anti-p38 (9212S) and anti-phospho-p38 (Thr180/Tyr182) (9211S) antibodies were purchased from Cell Signaling Technology (Danvers, MA, USA). The anti-PAD4 antibodies (17373-1-AP) were purchased from Proteintech Group, Inc. (Wuhan, HB, CHN). Horseradish peroxidase (HRP)-conjugated goat anti-mouse/rabbit IgG polymer kit was purchased from ZS GB-Bio (Beijing, CHN). Percoll™ PLUS (17-5445-01) was purchased from GE Healthcare (Uppsala, Sweden). The Cytometric Beads Array (CBA) kit (560485) was purchased from BD Biosciences (Becton, Dickinson and Company). Tris-buffered saline Tween-20 (TBST) was purchased from Biorigin (Beijing, CHN). Sodium citrate antigen retrieval solution and RIPA buffer were obtained from Solarbio (Beijing, CHN). The enhanced chemiluminescence (ECL) reagent was obtained from Cwbio IT Group (Beijing, CHN). Cell Counting Kit 8 (CCK-8) was purchased from Analysis Quiz (Beijing, CHN).
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Publication 2023
Alexa Fluor 555 Anti-Antibodies anti-IgG Antibodies Antigens BECN1 protein, human Buffers Chemiluminescence Freund's Adjuvant GAPDH protein, human Goat Horseradish Peroxidase Lipopolysaccharides Mitogen-Activated Protein Kinase 3 Mus Percoll Polymers Rabbits Radioimmunoprecipitation Assay Saline Solution sinomenine Sodium Citrate Tetradecanoylphorbol Acetate Tween 20
The human monocyte cell line THP-1 cells were purchased from ATCC. THP-1 cells were incubated in RPMI-1640 medium containing 10 mmol/L HEPES, 10% FBS and 1% penicillin/streptomycin antibiotic mixture, and then incubated for 24 h at 37 °C in a constant temperature incubator containing 5% CO2. Subsequently, in order to induce THP-1 cells to become adherent macrophages, the cells were seeded into 6-well plates, and the culture medium involving PMA (phorbol 12-myristate 13-acetate) at a final concentration of 100 ng/mL was added, and the cells was incubated for another 24 h. For cell model construction, the cells were cultured in serum-free medium containing different concentrations of oxidized low-density lipoprotein (ox-LDL) for 24 h based on the previously published methods [14 (link)].
For cell transfection, according to the instructions of the kit, miR-218-5p mimic and mimic-negative control (mimic-NC) were transfected into THP-1 cells using Lipofectamine 2000 transfection reagent. The transfection efficiency was detected by RT-qPCR 6 h after transfection.
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Publication 2023
Cell Lines Cells Culture Media HEPES Homo sapiens lipofectamine 2000 Macrophage Monocytes oxidized low density lipoprotein Penicillins Serum Streptomycin Tetradecanoylphorbol Acetate THP-1 Cells Transfection

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Publication 2023
Anti-Antibodies Antibodies, Anti-Idiotypic CD4 Positive T Lymphocytes Cells CY5.5 cyanine dye Cytokine Fluorescein-5-isothiocyanate Fluorescent Antibody Technique Helper-Inducer T-Lymphocyte IFNG protein, mouse IL2RA protein, human Ionomycin ITGAM protein, human Mus Proteins Protoplasm Regulatory T-Lymphocytes Tetradecanoylphorbol Acetate Th17 Cells Type-2 Helper T Cell

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Ionomycin is a laboratory reagent used in cell biology research. It functions as a calcium ionophore, facilitating the transport of calcium ions across cell membranes. Ionomycin is commonly used to study calcium-dependent signaling pathways and cellular processes.
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Phorbol 12-myristate 13-acetate is a laboratory compound used as a chemical tool in research. It functions as a potent activator of protein kinase C, a family of enzymes involved in various cellular processes.
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The PMA is a versatile laboratory equipment designed for precision measurement and analysis. It functions as a sensitive pressure transducer, accurately measuring and monitoring pressure levels in various applications. The PMA provides reliable and consistent data for research and testing purposes.
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Phorbol 12-myristate 13-acetate (PMA) is a chemical compound that acts as a potent protein kinase C activator. It is commonly used as a research tool in cell biology and biochemistry laboratories.
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Fetal Bovine Serum (FBS) is a cell culture supplement derived from the blood of bovine fetuses. FBS provides a source of proteins, growth factors, and other components that support the growth and maintenance of various cell types in in vitro cell culture applications.
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GolgiStop is a cell culture reagent that inhibits protein transport from the endoplasmic reticulum to the Golgi apparatus, thereby preventing the secretion of newly synthesized proteins. It is a useful tool for investigating protein trafficking and localization in cells.
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Brefeldin A is a fungal metabolite that inhibits the function of Golgi apparatus in eukaryotic cells. It acts by blocking the exchange of materials between the endoplasmic reticulum and Golgi compartments, leading to the collapse of the Golgi structure.
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GolgiPlug is a laboratory product designed to inhibit protein transport from the Golgi apparatus to the cell surface. It functions by blocking the secretory pathway, preventing the release of proteins from the Golgi complex. GolgiPlug is intended for use in cell biology research applications.
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RPMI 1640 medium is a commonly used cell culture medium developed at Roswell Park Memorial Institute. It is a balanced salt solution that provides essential nutrients, vitamins, and amino acids to support the growth and maintenance of a variety of cell types in vitro.
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RPMI 1640 is a common cell culture medium used for the in vitro cultivation of a variety of cells, including human and animal cells. It provides a balanced salt solution and a source of essential nutrients and growth factors to support cell growth and proliferation.

More about "Tetradecanoylphorbol Acetate"

Tetradecanoylphorbol acetate (TPA), also known as 12-O-Tetradecanoylphorbol-13-acetate (PMA), is a potent tumor-promoting agent and a widely used activator of protein kinase C (PKC).
It is frequently utilized in cell biology and cancer research to investigate signaling pathways, cell differentiation, and tumor progression.
PubCompare.ai can optimize your TPA/PMA research by leveraging AI to identify the best protocols from scientific literature, preprints, and patents, enhancing reproducibility and accuracy.
This tool can help you find the optimal products and procedures for your TPA/PMA experiments, taking your research to the next level.
Ionomycin is another commonly used reagent that works synergistically with TPA/PMA to activate signaling cascades and induce cellular responses.
The combination of TPA/PMA and Ionomycin is often employed to study immune cell activation, cytokine production, and cell differentiation.
Phorbol esters like TPA/PMA are structural analogs of diacylglycerol (DAG), a natural activator of PKC.
By binding to and activating PKC, TPA/PMA can mimic the effects of DAG and trigger a wide range of downstream signaling events.
When conducting TPA/PMA experiments, it is important to consider factors such as cell type, culture conditions (e.g., RPMI 1640 medium, FBS), and the use of inhibitors like GolgiStop (containing Brefeldin A) or GolgiPlug to study specific cellular processes.
These experimental parameters can greatly influence the observed cellular responses to TPA/PMA stimulation.