Background

12-O-tetradecanoyl phorbol-13-acetate (TPA) is an activator of ERK/MAPK with the concentration of 50¦Ìm [1].

ERK is an extracellular signal-regulated kinase and transfers signals from a receptor on the cell surface to DNA cooperated with MAPK. It is reported that ERK deficiency leads to the cell uncontrolled growth and is regarded as a target to cure cancers.

TPA is a potent ERK activator. When tested with A549 cells (human lung cancer cell line), TPA treatment led to anearly, strong, and relatively transient ERK phosphorylation [2]. In mouse embryo fibroblasts from DUSP5 (+/+) mice, administration of TPA increased levels of ERK expression [3].

Chemical Name

Canonical SMILES

N/A CCCCCCCCCCCCCC(=O)OC1C(C2(C(C=C(CC3(C2C=C(C3=O)C)O)CO)C4C1(C4(C)C)OC(=O)C)O)C

General tips For obtaining a higher solubility , please warm the tube at 37 ¡æ and shake it in the ultrasonic bath for a while.Stock solution can be stored below -20¡æ for several months.

Shipping Condition

Structure

Evaluation sample solution : ship with blue ice All other available size: ship with RT , or blue ice upon request.

Protocol

Kinase experiment [1]:

Protein kinase C was assayed by measuring the incorporation of 32P into H1 histone from [¦Ã-32P]ATP. The standard reaction mixture (0.25 ml) contained 5 pmol of Tris/HCl at pH 7.5,

1.25 pmol of magnesium nitrate, 50 pg of H1 histone, 2.5 nmol of [¦Ã-32P]ATP (5 to 15xl04 cpm/nmol), and 0.5 pg of protein kinase C. Phospholipid, diolein, phorbol esters, and Ca2+ were added as indicated in each experiment. All reagents were taken up in water which was

Kinase assays

Cell experiment [2]:

prepared by a double distillation apparatus followed by passing through a Chelex 100 column to remove as much Ca2+ as possible. After incubation for 3 min at 30oC, the reaction was stopped by the addition of 25% trichloroacetic acid, and acid-precipitable materials were collected on a Toyo-Roshi membrane filter (pore size, 0.45 pm). The catalytic fragment of protein kinase C was assayed similarly except that Ca2+, phospholipid, and diolein were omitted.

Cell lines B-lymphocyte cell line

The solubility of this compound in DMSO is >10 mM. General tips for obtaining a higher

Preparation method

concentration: Please warm the tube at 37¡æ for 10 minutes and/or shake it in the ultrasonic bath for a while. Stock solution can be stored below -20¡æ for several months.

Reacting condition 1 nM

Applications 12-O-tetradecanoyl phorbol-13-acetate was used for the activation of PKC (protein kinase C) in cells.

Animal experiment [3]:

Animal models Chemical skin carcinogenesis mice

Dosage form Twice weekly treatment (12.5 ¦Ìg in 100 ¦ÌL acetone)

Application 12-O-tetradecanoyl phorbol-13-acetate was used to induce skin cancer in mice.

Other notes Please test the solubility of all compounds indoor, and the actual solubility may slightly differ with the theoretical value. This is caused by an experimental system error and it is normal.

References:

[1] Castagna M, et al. Direct activation of calcium-activated, phospholipid-dependent protein kinase by tumor- promoting phorbol esters. J Biol Chem. 1982 Jul 10;257(13):7847-51.

[2] Jensen WA, et al. Inhibition of protein kinase C results in decreased expression of bovine leukemia virus. J Virol. 1992 Jul;66(7):4427-33.

[3] Rushworth LK, et al. Dual-specificity phosphatase 5 regulates nuclear ERK activity and suppresses skin cancer by inhibiting mutant Harvey-Ras (HRasQ61L)-driven SerpinB2 expression. Proc Natl Acad Sci U S A. 2014 Dec 23;111(51):18267-72.