PMA – Standard Reagent for THP1 Cell Differentiation

Phorbol 12-myristate 13-acetate (PMA), also known as 12-O-tetradecanoylphorbol-13-acetate or TPA, is a naturally occurring phorbol ester found in plants like croton oil.

PMA is commonly used to activate protein kinase C (PKC), a family of enzymes involved in various cellular processes such as cell growth, differentiation, proliferation and apoptosis (programmed cell death). PMA can activate all isoforms of PKC, but it has a particularly strong affinity for PKCα, PKCε and PKCδ. PMA-activated PKC can phosphorylate a variety of target proteins, including transcription factors, signalling molecules and cytoskeletal proteins. PKC is a critical component of many signalling pathways and its activation can lead to a cascade of intracellular events.

• Cell growth and differentiation: PMA can promote cell growth and differentiation by activating PKC. This effect is thought to be mediated by the activation of genes that promote cell growth and differentiation.
• Apoptosis: PMA can also induce apoptosis or programmed cell death. This effect is thought to be mediated by the activation of genes that promote apoptosis.
• Cancer: PMA is a known tumour promoter. This means that it can increase the risk of cancer by promoting cell growth and differentiation.
• Inflammation: PMA can induce inflammation. This effect is thought to be mediated by the activation of genes that promote inflammation.

Differentiation of THP-1 Cells

The human monocytic cell line, THP-1, is the most widely used cell line for in vitro studies investigating primary human macrophage function.  The reason is that following the differentiation of THP-1 cells using PMA, they acquire a macrophage-like phenotype, which mimics in many respects, primary human macrophages (M0 macrophages). PMA is a potent activator of protein kinase C (PKC), which is a key regulator of macrophage differentiation. When PMA is added to THP-1 cells, it causes them to express the surface markers CD14, CD16 and CD68, which are characteristic of M0 macrophages. PMA also induces the production of pro-inflammatory cytokines by M0 macrophages. Further treatment with PMA can activate M0 macrophages and differentiate them into M1 or M2 macrophages. 

Activation of M0 Macrophages

M0 macrophages are a type of macrophages that are not yet fully activated. They are characterised by their expression of a variety of surface markers, including CD14, CD16 and CD68. M0 macrophages are also able to produce a variety of cytokines, including IL-1β, IL-6 and TNF-α.

The addition of PMA M0 macrophages activates them through PKC activation. Consequently, activated M0 macrophages undergo a number of changes, including the increased expression of pro-inflammatory cytokines, increased production of reactive oxygen species, increased phagocytic activity and increased migration.

• PMA can induce the differentiation of M0 macrophages into M1 macrophages, which are pro-inflammatory cells that play a role in fighting infection.
• PMA can also induce the differentiation of M0 macrophages into M2 macrophages, which are anti-inflammatory cells that play a role in tissue repair.
• PMA can increase the production of reactive oxygen species by M0 macrophages, which can help to kill bacteria and other pathogens.
• PMA can increase the phagocytic activity of M0 macrophages, which allows them to engulf and destroy foreign particles.
• PMA can increase the migration of M0 macrophages, which allows them to travel to the site of infection or injury.

The differentiation of THP-1 cells into M0 macrophages is a complex process, important for the immune response to infection and injury.

Literature References:

1. In vitro studies on the mode of action of the phorbol esters, potent tumor promoters: part 1: P.M. Blumberg; Crit. Rev. Toxicol. 8, 153 (1980)
2. In vitro studies on the mode of action of the phorbol esters, potent tumor promoters: part 2: P.M. Blumberg; Crit. Rev. Toxicol. 8, 199 (1981)
3. The choice of phorbol 12-myristate 13-acetate differentiation protocol influences the response of THP-1 macrophages to a pro-inflammatory stimulus: M.E. Lund, et al.; J. Immunol. Methods 430, 64 (2016)
4. Standardized protocols for differentiation of THP-1 cells to macrophages with distinct M(IFNγ+LPS), M(IL-4) and M(IL-10) phenotypes: E.W. Baxter, et al.; J. Immunol. Methods 478, 112721 (2020)

Phorbol 12-myristate 13-acetate (PMA)

Other names: TPA; 12-O-Tetradecanoylphorbol 13-acetate

PMA is the most common phorbol. It is a potent tumour promoter and highly inflammatory in nature. It is a potent activator of protein kinase C (PKC), a family of enzymes that play a role in many cellular processes, including cell growth, differentiation and apoptosis. It is a standard reagent for THP1 cell differentiation.

AG-CN2-0010 (1 mg, 5 mg, 10 mg, 25 mg, BULK)

AdipoGen Life Sciences is an original Manufacturer of high-purity PMA. BULK quantities are available from Stock!

Small Molecule PKC Agonists/Inducers/Activators

Product Name	Product Code	Product Description
Ingenol-3-angelate (I3A)	AG-CN2-0012	Specific PKC activator.
Phorbol 12-myristate 13-acetate (PMA)	AG-CN2-0010	Potent PKC activator.

PKC Antagonist/Inhibitor/Blocker

Product Name	Product Code	Product Description
Staurosporine	AG-CN2-0022	Potent, cell-permeable, reversible, ATP-competitive broad spectrum PKC antagonist.
Bisindolylmaleimide I	AG-CR1-0009	Cell-permeable, selective PKC inhibitor (Ki = 10 nM).
Bisindolylmaleimide I . hydrochloride	AG-CR1-0110	Cell-permeable, selective PKC inhibitor (Ki = 10 nM), water-soluble.
Bisindolylmaleimide II	AG-CR1-0010	PKC antagonist.
Bisindolylmaleimide III	AG-CR1-0112	Potent and selective PKC inhibitor.
Bisindolylmaleimide IV	AG-CR1-0152	Cell-permeable PKC inhibitor.
Bisindolylmaleimide VIII . acetate	AG-CR1-0114	Selective PKC inhibitor.
Bisindolylmaleimide IX . methanesulfonate	AG-CR1-0111	Selective, cell-permeable PKC inhibitor.
Bisindolylmaleimide X . hydrochloride	AG-CR1-0113	Selective PKC inhibitor.
Bisindolylmaleimide XI . hydrochloride	AG-CR1-0109	Selective cell-permeable PKC inhibitor.
Calphostin C	AG-CN2-0430	Potent, highly specific, cell-permeable, light-dependent PKC antagonist.

Negative Controls for PKC Modulators

Product Name	Product Code	Product Description
Bisindolylmaleimide V	AG-CR1-0023	Negative control for PKC inhibitors.
4α-Phorbol 12-myristate 13-acetate	AG-CN2-0082	Negative control for phorbol ester activation of PKC and studies with PMA.

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Originally posted by Adipogen on: https://adipogen.com/phorbol-12-myristate-13-acetate-pma

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