佛波醇12-十四酸酯13-乙酸酯作用機制 - Medchemexpress - MCE中國

1、Product Data SheetPhorbol 12-myristate 13-acetateCat. No.: HY-18739CAS No.: 16561-29-8分式: CHO分量: 616.83作靶點: PKC; SPHK作通路: Epigenetics; TGF-beta/Smad; Immunology/Inflammation儲存式: 4C, protect from light* In solvent : -80C, 6 months; -20C, 1 month (protect from light)溶解性數(shù)據(jù)體外實驗 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制備儲備液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請根據(jù)產(chǎn)品在不同溶劑中的溶解度選擇合適的溶劑配制儲備液；旦配成溶液，請分

3、裝保存，避免反復凍融造成的產(chǎn)品失效。儲備液的保存式和期限：-80C, 6 months; -20C, 1 month (protect from light)。-80C 儲存時，請在 6 個內(nèi)使，-20C 儲存時，請在 1 個內(nèi)使。體內(nèi)實驗請根據(jù)您的實驗動物和給藥式選擇適當?shù)娜芙獍?。以下溶解案都請先按?In Vitro 式配制澄清的儲備液，再依次添加助溶劑：為保證實驗結(jié)果的可靠性，澄 的儲備液可以根據(jù)儲存條件，適當保存；體內(nèi)實驗的作液，建議您現(xiàn)現(xiàn)配，當天使； 以下溶劑前顯的百分 指該溶劑在您配制終溶液中的體積占；如在配制過程中出現(xiàn)沉淀、析出現(xiàn)象，可以通過加熱和/或超聲的式助溶1. 請依序添加

4、每種溶劑： 10% DMSO 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可獲得 2.5 mg/mL (4.05 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 400 L PEG300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。2. 請依序添加每種溶劑： 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此案可獲得 2.5 mg/mL (4.05 mM) 的均勻懸濁液，懸濁液可于服和腹腔注射。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。3. 請依序添加每種溶劑： 10% DMSO 90% corn oilSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可獲得 2.5 m

6、g/mL (4.05 mM，飽和度未知) 的澄 溶液，此案不適于實驗周 期在半個以上的實驗。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 DMSO 儲備液加到 900 L 油中，混合均勻。4. 請依序添加每種溶劑： 10% EtOH 40% PEG300 5% Tween-80 45% salineSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可獲得 2.5 mg/mL (4.05 mM，飽和度未知) 的澄清溶液。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 EtOH 儲備液加到 400 L PEG

7、300 中，混合均勻；向上述體系中加50 L Tween-80，混合均勻；然后繼續(xù)加 450 L 理鹽定容 1 mL。5. 請依序添加每種溶劑： 10% EtOH 90% (20% SBE-CD in saline)Solubility: 2.5 mg/mL (4.05 mM); Suspended solution; Need ultrasonic此案可獲得 2.5 mg/mL (4.05 mM) 的均勻懸濁液，懸濁液可于服和腹腔注射。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 EtOH 儲備液加到 900 L 20% 的 SBE-CD 理鹽溶液中，混合均勻。6. 請

8、依序添加每種溶劑： 10% EtOH 90% corn oilSolubility: 2.5 mg/mL (4.05 mM); Clear solution此案可獲得 2.5 mg/mL (4.05 mM，飽和度未知) 的澄 溶液，此案不適于實驗周 期在半個以上的實驗。以 1 mL 作液為例，取 100 L 25.0 mg/mL 的澄 EtOH 儲備液加到 900 L 油中，混合均勻。BIOLOGICAL ACTIVITY物活性 Phorbol 12-myristate 13-acetate (PMA)種佛波酯， 蛋激酶 C (PKC) 和 SphK 的激活劑。IC & Target PKC1

9、1.7 nM (EC50)體外研究 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.體內(nèi)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.戶使本產(chǎn)品發(fā)表的科研獻 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