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Phorbol 12-myristate 13-acetate (PMA; TPA; Phorbol myristate acetate), a phorbol ester, is a dual SphK and protein kinase C (PKC) activator. Phorbol 12-myristate 13-acetate is a NF-κB activator. Phorbol 12-myristate 13-acetate induces differentiation in THP-1 cells (Validated by MedChemExpress (MCE)).
The migratory and invasive capabilities of HOS and MG-63 cells were assessed using Transwell assays in response to normoxic or hypoxic M0, M1, and M2 macrophages. THP-1 cells underwent differentiation into M0 macrophages following 24 h exposure to 100 ng/mL phorbol 12-myristate 13-acetate (PMA, HY-18739). Following a 24 h recovery in a fresh complete medium, cells were treated with 100 ng/mL lipopolysaccharide and 20 ng/mL IFN-gamma Protein, Human (IFN-γ, HY-P7025A) for 48 h to drive M1 polarization.
Western Blot (WB) analysis ofADAM22 expression in indicated cells after PMA (150 nM) or forskolin (50 μM) administration for 0 hour, 24 hours, 48 hours, and 72 hours.
Total lysates from cells are analyzed for the expression of MMP-2 and MMP-9 by Western blot analysis in the treatment of different concentrations of PMA and PDD.
MDA-MB-231 cells are pretreated with PPD for 24 h followed by exposure to 50 ng/mL of PMA for 30 min. The whole cell lysates are analyzed by Western blot for the activation of MAPK.
Bar graphs show levels of GABAAR-α2 mRNA and GABAAR-α2 protein in BLA of control mice treated with BLA-injection of H89 or GF109203X (GFX); in MPTP-mice treated with BLA-injection of PMA, or the co-administration of quinpirole and H89 (quin/+H89) or GF109203X (quin/+GFX).
Phorbol 12-myristate 13-acetate (PMA; TPA; Phorbol myristate acetate), a phorbol ester, is a dual SphK and protein kinase C (PKC) activator[1][2]. Phorbol 12-myristate 13-acetate is a NF-κB activator. Phorbol 12-myristate 13-acetate induces differentiation in THP-1 cells (Validated by MedChemExpress (MCE))[3][7].
Antiviral activity against Chikungunya virus 899 infected in african green monkey Vero cells assessed as inhibition of virus-induced cytopathic effect incubated for 6 to 7 days by MTS assay
Antiviral activity against Chikungunya virus 899 infected in african green monkey Vero cells assessed as inhibition of virus-induced cytopathic effect incubated for 6 to 7 days by MTS assay
Antiviral activity against Sindbis virus HRsp infected in african green monkey Vero cells assessed as inhibition of virus-induced cytopathic effect incubated for 6 to 7 days by MTS assay
Antiviral activity against Sindbis virus HRsp infected in african green monkey Vero cells assessed as inhibition of virus-induced cytopathic effect incubated for 6 to 7 days by MTS assay
Antiviral activity against chikungunya virus Indian ocean strain 899 infected in Vero cells assessed as virus-induced cytopathic effect after 6 to 7 days
Antiviral activity against chikungunya virus Indian ocean strain 899 infected in Vero cells assessed as virus-induced cytopathic effect after 6 to 7 days
Antiviral activity against Semliki forest virus infected in african green monkey Vero cells assessed as inhibition of virus-induced cytopathic effect incubated for 6 to 7 days by MTS assay
Antiviral activity against Semliki forest virus infected in african green monkey Vero cells assessed as inhibition of virus-induced cytopathic effect incubated for 6 to 7 days by MTS assay
PMA (100, 200 ng/mL; 1-5 days) induce THP-1 cells to differentiate into macrophage-like cells (THP-1 macrophages), characterized by changes in morphology (adherent macrophage-like phenotype), and increases cell surface expression of CD11b[3][5].
PMA (20 ng/mL, 36 h) inhibits endothelial cell migration through activating the PKC-δ/Syk/NF-κB-mediated up-regulation of Thy-1[7].
MedChemExpress (MCE) has not independently confirmed the accuracy of these methods. They are for reference only.
Phorbol 12-myristate 13-acetate 関連抗体
体内実験
Note:
Please do not refer to only one article to determine the experimental conditions. It is recommended to determine the optimal experimental conditions (animal strain, age, dosage, frequency and cycle, detection time and indicators, etc.) through preliminary experiments before the formal experiment.
Phorbol 12-myristate 13-acetate (PMA) can be used to create models of ear edema[8][9].
PMA induces a pronounced inflammatory response mediated by protein kinase C (PKC), specifically activating PLA2 to trigger inflammation.
Specific Modeling Methods
Mice: Swiss mouse • Female • 25-30 g Administration: Topically applied in one ear • 100 μg/mL in 20 μL (2 μg/ear) vehicle • single dose
Note
Inhibitory or reventable compounds' administration should be performed before modeling. Such as Hydroxyachillin, should be administered 1 h before PMA treatment.
Modeling Indicators
Appearance monitoring: The thickness difference between the left and right ears increases significantly. Indicator changes: Increased vascular permeability.
PMA induces a pronounced inflammatory response mediated by protein kinase C (PKC), specifically activating PLA2 to trigger inflammation.
Specific Modeling Methods
Rat: Wistar • male • adult with weight of 200-220 g Mice: Swiss albino • male • 25-30 g Administration: Topically applied in one ear • 2.5 μg in 20 μL vehicle • single dose
Note
Inhibitory compounds' administration should be conducted 4 h before mouse were killed.
Modeling Indicators
Appearance monitoring: The quality difference between the left and right ears increases significantly. Indicator changes: Stimulate macrophages to produce superoxide anions.
DMSO : 25 mg/mL (40.53 mM; Need ultrasonic; Hygroscopic DMSO has a significant impact on the solubility of product, please use newly opened DMSO)
Preparing Stock Solutions
ConcentrationSolventMass
1 mg
5 mg
10 mg
1 mM
1.6212 mL
8.1060 mL
16.2119 mL
5 mM
0.3242 mL
1.6212 mL
3.2424 mL
10 mM
0.1621 mL
0.8106 mL
1.6212 mL
View the Complete Stock Solution Preparation Table
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
For the following dissolution methods, please ensure to first prepare a clear stock solution using an In Vitro approach and then sequentially add co-solvents:
To ensure reliable experimental results, the clarified stock solution can be appropriately stored based on storage conditions. As for the working solution for in vivo experiments, it is recommended to prepare freshly and use it on the same day. The percentages shown for the solvents indicate their volumetric ratio in the final prepared solution. If precipitation or phase separation occurs during preparation, heat and/or sonication can be used to aid dissolution.
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 2
Add each solvent one by one: 10% DMSO 90% (20% SBE-β-CD in Saline)
Solubility: 2.5 mg/mL (4.05 mM); Suspended solution; Need ultrasonic
This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 3
Add each solvent one by one: 10% DMSO 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (4.05 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μLDMSO stock solution (25.0 mg/mL) to 900 μLCorn oil, and mix evenly.
Protocol 4
Add each solvent one by one: 10% EtOH 40% PEG300 5% Tween-80 45% Saline
Solubility: ≥ 2.5 mg/mL (4.05 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown).
Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 400 μL PEG300, and mix evenly; then add 50 μL Tween-80 and mix evenly; then add 450 μL Saline to adjust the volume to 1 mL.
Preparation of Saline: Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution.
Protocol 5
Add each solvent one by one: 10% EtOH 90% (20% SBE-β-CD in Saline)
Solubility: 2.5 mg/mL (4.05 mM); Suspended solution; Need ultrasonic
This protocol yields a suspended solution of 2.5 mg/mL. Suspended solution can be used for oral and intraperitoneal injection.
Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 900 μL 20% SBE-β-CD in Saline, and mix evenly.
Preparation of 20% SBE-β-CD in Saline (4°C, storage for one week): 2 g SBE-β-CD powder is dissolved in 10 mL Saline, completely dissolve until clear.
Protocol 6
Add each solvent one by one: 10% EtOH 90% Corn Oil
Solubility: ≥ 2.5 mg/mL (4.05 mM); Clear solution
This protocol yields a clear solution of ≥ 2.5 mg/mL (saturation unknown). If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Taking 1 mL working solution as an example, add 100 μL EtOH stock solution (25.0 mg/mL) to 900 μLCorn oil, and mix evenly.
In Vivo Dissolution Calculator
Please enter the basic information of animal experiments:
Dosage
mg/kg
Animal weight (per animal)
g
Dosing volume (per animal)
μL
Number of animals
Recommended: Prepare an additional quantity of animals to account for potential losses during experiments.
Please enter your animal formula composition:
%
DMSO+
%
+
%
Tween-80
+
%
Saline
Recommended: Keep the proportion of DMSO in working solution below 2% if your animal is weak.
The co-solvents required include: DMSO,
. All of co-solvents are available by MedChemExpress (MCE).
, Tween 80. All of co-solvents are available by MedChemExpress (MCE).
Calculation results:
Working solution concentration:
mg/mL
Method for preparing stock solution:
mg
drug dissolved in
μL
DMSO (Stock solution concentration: mg/mL).
The concentration of the stock solution you require exceeds the measured solubility. The following solution is for reference only. If necessary, please contact MedChemExpress (MCE).
Method for preparing in vivo working solution for animal experiments: Take
μL DMSO stock solution, add
μL .
μL , mix evenly, next add
μL Tween 80, mix evenly, then add
μL Saline.
Dissolve 0.9 g sodium chloride in ddH₂O and dilute to 100 mL to obtain a clear Saline solution
If the continuous dosing period exceeds half a month, please choose this protocol carefully.
Please ensure that the stock solution in the first step is dissolved to a clear state, and add co-solvents in sequence. You can use ultrasonic heating (ultrasonic cleaner, recommended frequency 20-40 kHz), vortexing, etc. to assist dissolution.
αT3-1 and LβT-2 cells are grown in monolayer cultured in DMEM in humidified incubator 5% CO2 at 37°C. Serum starvation is with 0.1% FCS in the same medium for 16 h. GnRH and PMA are then added for the length of time as indicated. In general, αT3-1 cells are transiently transfected by ExGen 500 or by jetPRIME, while LβT2 cells only by jetPRIME transfection reagent. For experiments with dominant-negative (DN) PKCs, αT3-1 cells (in 6 cm plates) are transfected with 1.5 μg of p38α-GFP with 3 μg of control vector, pCDNA3, or with 3 μg of the DN-PKCs constructs. For LβT2 cells, transfections are performed (in 10 cm plates) with 4 μg of p38α-GFP along with 9 μg of control vector, pCDNA3, or with 9 μg of the DN-PKCs constructs. Approximately 30 h after transfection, the cells are serum starved (0.1% FCS) for 16 h and later stimulated with GnRH or PMA, washed twice with ice-cold PBS, treated with the lysis buffer, followed by one freeze-thaw cycle. Cells are harvested; following centrifugation (15,000×g, 15 min, 4°C) supernatants are taken for immunoprecipitation experiments[2].
Rats[3] All experiments qre performed with male Wistar rats (weighing 250-280 g). One hundred and thirty-five Wistar rats are randomly divided into seven groups. (1) Rats in the sham group (n=21) are given a lateral cerebral ventricle injection of 0.9% normal saline; (2) Rats in the IR group (n=21) are given a lateral cerebral ventricle injection of 0.9% normal saline 30 min before middle cerebral artery occlusion (MCAO); (3) Rats in the Carbenoxolone (CBX) group (n=21) are given a lateral cerebral ventricle injection of CBX (5 μg/mL×10 μL) 30 min before MCAO; (4) Rats in the Sch-6783 group (n=21) are given a lateral cerebral ventricle injection of DZX (2 mM×30 μL) 30 min prior to MCAO; (5) Rats in the 5-HD group (n=21) are given a lateral cerebral ventricle injection of 5-HD (100 mM×10 μL), and after 10 min, DZX is injected 15 min prior to MCAO; (6) The rats in the DZX + Ro group (n=15) are given a lateral cerebral ventricle injection of DZX, and after 10 min, Ro-31-8425 (400 μg/kg) is injected 15 min prior to MCAO; (7) The rats in the 5-HD+PMA group (n=15) are given an intraperitoneal injection of PMA (200 μg/kg) after the injection of 5-HD and DZX.
*Please refer to the solubility information to select the appropriate solvent. Once prepared, please aliquot and store the solution to prevent product inactivation from repeated freeze-thaw cycles. Storage method and period of stock solution: -80°C, 6 months; -20°C, 1 month (protect from light). When stored at -80°C, please use it within 6 months. When stored at -20°C, please use it within 1 month.
Optional Solvent
ConcentrationSolventMass
1 mg
5 mg
10 mg
25 mg
DMSO / Ethanol
1 mM
1.6212 mL
8.1060 mL
16.2119 mL
40.5298 mL
5 mM
0.3242 mL
1.6212 mL
3.2424 mL
8.1060 mL
10 mM
0.1621 mL
0.8106 mL
1.6212 mL
4.0530 mL
15 mM
0.1081 mL
0.5404 mL
1.0808 mL
2.7020 mL
20 mM
0.0811 mL
0.4053 mL
0.8106 mL
2.0265 mL
25 mM
0.0648 mL
0.3242 mL
0.6485 mL
1.6212 mL
30 mM
0.0540 mL
0.2702 mL
0.5404 mL
1.3510 mL
40 mM
0.0405 mL
0.2026 mL
0.4053 mL
1.0132 mL
Ethanol
50 mM
0.0324 mL
0.1621 mL
0.3242 mL
0.8106 mL
60 mM
0.0270 mL
0.1351 mL
0.2702 mL
0.6755 mL
80 mM
0.0203 mL
0.1013 mL
0.2026 mL
0.5066 mL
100 mM
0.0162 mL
0.0811 mL
0.1621 mL
0.4053 mL
Phorbol 12-myristate 13-acetate Related Classifications
Species cross-reactivity must be investigated individually for each product. Many human cytokines will produce a nice response in mouse cell lines, and many mouse proteins will show activity on human cells. Other proteins may have a lower specific activity when used in the opposite species.
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バルクお問い合わせ
Inquiry Information
製品名:
Phorbol 12-myristate 13-acetate
製品番号:
HY-18739
数量:
MCE 日本正規代理店:
カスタマーレビュー
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1/16
1/16
MedChemExpress Validation THP-1 cells were seeded at 2 × 105 cells/well in a 24 well plate and treated with 100 and 200 ng/mL Phorbol 12-myristate 13-acetate (PMA) (HY-18739) for 24 or 48 h. The results indicated that PMA induces THP-1 cell differentiation.
1/16
MedChemExpress Validation THP-1 cells were seeded at 2 × 105 cells/well in a 24 well plate and treated with 100 and 200 ng/mL Phorbol 12-myristate 13-acetate (PMA) (HY-18739) for 24 h. Cells were lysed and the expression of CD11b was measured. The results indicated that PMA increases expression of CD11b.
1/16
MedChemExpress Validation THP-1 cells were seeded at 3 × 105 cells/well in a 12 well plate and treated with 100 and 200 ng/mL Phorbol 12-myristate 13-acetate (PMA) (HY-18739) for 24 h. The results indicated that PMA induces THP-1 cell differentiation and showed lot-to-lot consistency.
Customer Validation
The migratory and invasive capabilities of HOS and MG-63 cells were assessed using Transwell assays in response to normoxic or hypoxic M0, M1, and M2 macrophages. THP-1 cells underwent differentiation into M0 macrophages following 24 h exposure to 100 ng/mL phorbol 12-myristate 13-acetate (PMA, HY-18739). Following a 24 h recovery in a fresh complete medium, cells were treated with 100 ng/mL lipopolysaccharide and 20 ng/mL IFN-gamma Protein, Human (IFN-γ, HY-P7025A) for 48 h to drive M1 polarization.
Customer Validation
Western Blot (WB) analysis ofADAM22 expression in indicated cells after PMA (150 nM) or forskolin (50 μM) administration for 0 hour, 24 hours, 48 hours, and 72 hours.
Customer Validation
Total lysates from cells are analyzed for the expression of MMP-2 and MMP-9 by Western blot analysis in the treatment of different concentrations of PMA and PDD.
Customer Validation
MDA-MB-231 cells are pretreated with PPD for 24 h followed by exposure to 50 ng/mL of PMA for 30 min. The whole cell lysates are analyzed by Western blot for the activation of MAPK.
Customer Validation
Bar graphs show levels of GABAAR-α2 mRNA and GABAAR-α2 protein in BLA of control mice treated with BLA-injection of H89 or GF109203X (GFX); in MPTP-mice treated with BLA-injection of PMA, or the co-administration of quinpirole and H89 (quin/+H89) or GF109203X (quin/+GFX).
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