佛波醇12-十四酸酯13-乙酸酯作用机制 - Medchemexpress - MCE中国

1、Product Data SheetPhorbol 12-myristate 13-acetateCat. No.: HY-18739CAS No.: 16561-29-8分式: CHO分量: 616.83作靶点: PKC; SPHK作通路: Epigenetics; TGF-beta/Smad; Immunology/Inflammation储存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性数据体外实验 Ethanol : 100 mg/mL (162.

2、12 mM; Need ultrasonic)DMSO : 50 mg/mL (81.06 mM)H2O : 0.1 mg/mL (insoluble)* means soluble, but saturation unknown.SolventMass1 mg 5 mg 10 mgConcentration制备储备液1 mM 1.6212 mL 8.1060 mL 16.2119 mL5 mM 0.3242 mL 1.6212 mL 3.2424 mL10 mM 0.1621 mL 0.8106 mL 1.6212 mL请根据产品在不同溶剂中的溶解度选择合适的溶剂配制储备液；旦配成溶液，请分

3、装保存，避免反复冻融造成的产品失效。储备液的保存式和期限：-80C, 6 months; -20C, 1 month (protect from light)。-80C 储存时，请在 6 个内使，-20C 储存时，请在 1 个内使。体内实验请根据您的实验动物和给药式选择适当的溶解案。以下溶解案都请先按照 In Vitro 式配制澄清的储备液，再依次添加助溶剂：为保证实验结果的可靠性，澄 的储备液可以根据储存条件，适当保存；体内实验的作液，建议您现现配，当天使； 以下溶剂前显的百分 指该溶剂在您配制终溶液中的体积占；如在配制过程中出现沉淀、析出现象，可以通过加热和/或超声的式助溶1. 请依序添加

4、每种溶剂： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可获得 2.5 mg/mL (4.05 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 400 L PEG300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。2. 请依序添加每种溶剂： 10% DMSO 90% (20% SBE-CD in saline)Solub

5、ility: 2.5 mg/mL (4.05 mM); Suspended solution; Need ultrasonicPage 1 of 2 www.MedChemE此案可获得 2.5 mg/mL (4.05 mM) 的均匀悬浊液，悬浊液可于服和腹腔注射。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。3. 请依序添加每种溶剂： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可获得 2.5 m

6、g/mL (4.05 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 DMSO 储备液加到 900 L 油中，混合均匀。4. 请依序添加每种溶剂： 10% EtOH 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可获得 2.5 mg/mL (4.05 mM，饱和度未知) 的澄清溶液。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 EtOH 储备液加到 400 L PEG

7、300 中，混合均匀；向上述体系中加50 L Tween-80，混合均匀；然后继续加 450 L 理盐定容 1 mL。5. 请依序添加每种溶剂： 10% EtOH 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (4.05 mM); Suspended solution; Need ultrasonic此案可获得 2.5 mg/mL (4.05 mM) 的均匀悬浊液，悬浊液可于服和腹腔注射。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 EtOH 储备液加到 900 L 20% 的 SBE-CD 理盐溶液中，混合均匀。6. 请

8、依序添加每种溶剂： 10% EtOH 90% corn oilSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可获得 2.5 mg/mL (4.05 mM，饱和度未知) 的澄 溶液，此案不适于实验周 期在半个以上的实验。以 1 mL 作液为例，取 100 L 25.0 mg/mL 的澄 EtOH 储备液加到 900 L 油中，混合均匀。BIOLOGICAL ACTIVITY物活性 Phorbol 12-myristate 13-acetate (PMA)种佛波酯， 蛋激酶 C (PKC) 和 SphK 的激活剂。IC & Target PKC1

9、1.7 nM (EC50)体外研究 In order to examine the role of PKC in p38MAPK phosphorylation, the cells are stimulated with the PKC activator,PMA (100 nM), which mimics the binding of DAG, the natural activator of PKC, to the C1 region of the PKCs.p38MAPK phosphorylation by PMA is observed in the two cell types

10、 similar to that observed by GnRH in T3-1 cells,that is, a slow sustained activation (3.2-fold and 3.6-fold, respectively at 30 min). The paradoxical findings that PKCsactivated by GnRH and PMA play a differential role in p38MAPK phosphorylation may be explained by differentiallocalization of the PK

11、Cs. Basal, GnRH- and PMA- stimulation of p38MAPK phosphorylation in T3-1 cells is mediatedby Ca2+ influx via voltage-gated Ca2+ channels and Ca2+ mobilization, while in the differentiated LT2 gonadotropecells it is mediated only by Ca2+ mobilization2.体内研究 PMA is a PKC agonist, which reverses the dam

12、age induced by 5-hydroxydecanoic acid (5-HD). Thus, activation of themitoKATP protected mitochondrial function in SOD and MDA via the PKC pathway3.PROTOCOLCell Assay 2 T3-1 and LT-2 cells are grown in monolayer cultured in DMEM in humidified incubator 5% CO2 at 37C. Serumstarvation is with 0.1% FCS

13、in the same medium for 16 h. GnRH and PMA are then added for the length of time asPage 2 of 3 www.MedChemEindicated. In general, T3-1 cells are transiently transfected by ExGen 500 or by jetPRIME, while LT2 cells only byjetPRIME transfection reagent. For experiments with dominant-negative (DN) PKCs,

14、 T3-1 cells (in 6 cm plates) aretransfected with 1.5 g of p38-GFP with 3 g of control vector, pCDNA3, or with 3 g of the DN-PKCs constructs.For LT2 cells, transfections are performed (in 10 cm plates) with 4 g of p38-GFP along with 9 g of control vector,pCDNA3, or with 9 g of the DN-PKCs constructs.

15、 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 lysisbuffer, followed by one freeze-thaw cycle. Cells are harvested; following centrifugation (15,000g, 15 min, 4C)supernatan

16、ts are taken for immunoprecipitation experiments2.MCE has not independently confirmed the accuracy of these methods. They are for reference only.Animal Rats3Administration 3 All experiments qre performed with male Wistar rats (weighing 250-280 g). One hundred and thirty-five Wistar ratsare randomly

17、divided into seven groups. (1) Rats in the sham group (n=21) are given a lateral cerebral ventricleinjection 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

18、 the Carbenoxolone (CBX) group(n=21) are given a lateral cerebral ventricle injection of CBX (5 g/mL10 L) 30 min before MCAO; (4) Rats in theSch-6783 group (n=21) are given a lateral cerebral ventricle injection of DZX (2 mM30 L) 30 min prior to MCAO;(5) Rats in the 5-HD group (n=21) are given a lat

19、eral cerebral ventricle injection of 5-HD (100 mM10 L), and after10 min, DZX is injected 15 min prior to MCAO; (6) The rats in the DZX + Ro group (n=15) are given a lateral cerebralventricle injection of DZX, and after 10 min, Ro-31-8425 (400 g/kg) is injected 15 min prior to MCAO; (7) The rats inth

20、e 5-HD+PMA group (n=15) are given an intraperitoneal injection of PMA (200 g/kg) after the injection of 5-HDand DZX.MCE has not independently confirmed the accuracy of these methods. They are for reference only.户使本产品发表的科研献 Biomaterials. 2018 Jul;170:70-81. J Control Release. 2020 Jan 28;320:304-313.

21、 Acta Biomater. 2019 Apr 1;88:392-405. Cell Death Dis. 2020 Jan 30;11(1):78. Cell Death Dis. 2018 Aug 29;9(9):880.See more customer validations on HYPERLINK www.MedChemE www.MedChemEREFERENCES1. Sergio E. Alvarez, et al. Autocrine and paracrine roles of sphingosine-1-phosphate. TRENDS in Endocrinology and Metabolism Vol.18 No.82. Mugami S, et al. Differential roles of PKC isoforms (PKCs) and Ca2+ in GnRH and phorbol 12-myristate 13-acetate (PMA) stimulation of p38MAPKphosphorylation in immortalized gonadot