Tetradecanoylphorbol Acetate
It is frequently utilized in cell biology and cancer research to investigate signaling pathways, cell differentiation, and tumor progression.
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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.
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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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.