PBFI AM ( K+ Indicator) 鉀離子指示探針

【溫馨提示】：見(jiàn)我司整理的鉀離子載體探針（Potassium ionophoresindicators）產(chǎn)品專(zhuān)題。

產(chǎn)品簡(jiǎn)介：

PBFI，英文全名Potassium-binding Benzofuran Isophthalate，一種K+敏感的熒光探針，用來(lái)測定細胞和細胞內區隔（Intracellular compartments）的K+水平變化。雖然PBFI對K+的選擇能力弱于Ca2+指示劑比如Fura-2，但在其他一價(jià)陽(yáng)離子存在體系中，PBFI足以檢測K+的生理濃度。結合離子后的PBFI光譜變化可通過(guò)激發(fā)光比率測定來(lái)分析，其能與使用相同光濾片和儀器檢測的探針Fura-2共同使用。

PBFI對K+的解離常數（Kd）非常依賴(lài)于Na+的存在與否。在不含Na+的體系，PBFI對K+的Kd值為5.1mM；而在含135mM K+/ Na+總濃度（約為生理離子強度）的溶液體系，PBFI對K+的Kd值為44mM；若緩沖液中的Na+用四甲基氯化銨所替代，PBFI對K+的Kd值變?yōu)?span>11mM。氯化膽堿和N-甲基葡萄糖胺是培養基中另兩種可能替代Na+的化合物。雖然PBFI對K+的選擇性比Na+僅強1.5倍，這一選擇能力已足以滿(mǎn)足檢測需求，因為正常情況細胞內K+濃度比Na+高10倍左右。

本品為乙酰氧基甲基酯（Acetoxymethyl ester, AM ester）形式的PBFI，CAS NO：124549-23-1，具有細胞膜滲透性，只需簡(jiǎn)單孵育即可進(jìn)入細胞，常用加載濃度范圍5-10μM，加載時(shí)間40min-4h，根據具體的實(shí)驗要求和細胞類(lèi)型來(lái)調整。

產(chǎn)品特性：

1）化學(xué)名4,4'-[1,4,10,13-tetraoxa-7,16-diazacyclooctadecane-7,16-diylbis(5-methoxy-6,2-benzofurandiyl)]bis- 1,3-benzenedicarboxylic acid, tetrakis[(acetyloxy)methyl] ester

2）同義名：Potassium-binding Benzofuran Isophthalate Acetoxymethyl ester

3）CAS NO：124549-23-1

4）分子式：C58H62N2O24

5）分子量：1171.1

6）純度：≥95%

7）Ex/Em：~340,380/500 nm

8）外觀(guān)：黃色至橙色粉末

9）溶解性：溶于DMSO（10mM）和甲醇

保存條件: 

-20oC避光干燥保存，2年有效。

注意事項：

1.       PBFI AM易受潮，粉末需干燥保存；粉末需用無(wú)水DMSO溶解，配制儲存液（如10mM），置于-20℃干燥避光保存，小量分裝避免反復凍融，至少3個(gè)月穩定。

2.      PBFI AM由于水溶性較差，建議使用Pluronic F-127以?xún)?yōu)化探針的細胞加載效率。通常情況，將PBFI AM的DMSO儲存液與等體積Pluronic F-127（25% w/v）混合均勻，之后即刻加入適量的細胞加載緩沖液（cell loading buffer）中達到所需濃度。

3.      為了您的安全和健康，請穿實(shí)驗服并戴一次性手套操作。

文獻引用：

[1] Tong W et al. Phthalocyanine functionalized poly(glycidyl methacrylate) nano-assemblies for photodynamic inactivation of bacteria. Biomater. Sci., 2019,7, 1905-1918

[2] Li R et al. Biofilm inhibition and mode of action of epigallocatechin gallate against Vibrio mimicus. Food Control, Volume 113, July 2020, 107148

Then PBFI probe was added and incubated at 37 °C for 90 min. The cells were washed, collected and resuspended with PBS buffer. Aliquots (100 μL) of bacterial suspension were transferred to a Corning 96 well black plate, and 100 μL of various concentrations of EGCG were dispensed in the microtiter plate wells.

[3] Liu Y, Zhen W, Wang Y, Song S, Zhang H. Na2S2O8 Nanoparticles Trigger Antitumor Immunotherapy through Reactive Oxygen Species Storm and Surge of Tumor Osmolarity. J Am Chem Soc. 2020 Dec 30;142(52):21751-21757. doi: 10.1021/jacs.0c09482. Epub 2020 Dec 18. PMID: 33337859.

 4T1 cells were inoculated into glass bottom culture dishes for 24 h. Then, adding PNSO NPs medium solution (80 μg/mL) to continue co-culture for 4 h. The treated 4T1 cells were further incubated with 10μM Na+ indicator SBFI AM in 0.04% Pluronic F-127 and the fluorescence signal was measured by CLSM.

[4] Liang Z, Yang Y, Yu G, et al. Engineering aluminum hydroxyphosphate nanoparticles with well-controlled surface property to enhance humoral immune responses as vaccine adjuvants. Biomaterials. 2021 Jun;275:120960. DOI: 10.1016/j.biomaterials.2021.120960.

[5] BMDMs were treated with AAHPs (250 μg/mL) in the presence of LPS at 500 ng/mL for 5 h. Then PBFI AM was added to the cells at the concentration of 10 μM, and cells were incubated at 37 °C for 1h. Triton X-100 (0.2%) treated cells were used as controls. The fluorescence of PBFI AM was measured at the Ex/Em of 340/615 nm. The data were expressed as relative fluorescence intensity (RFI) defined as the fluorescence intensity of AAHPs-treated BMDMs normalized to the intensity of control cells.

[6] Jia Y, Yang B, Shi J, Fang D, Wang Z, Liu Y. Melatonin prevents conjugative transfer of plasmid-mediated antibiotic resistance genes by disrupting proton motive force. Pharmacol Res. 2022 Jan;175:105978. doi: 10.1016/j.phrs.2021.105978. Epub 2021 Nov 21. PMID: 34813930.

As for the detection of intracellular K + concentration, the PBFI-AM (K + indicator) fluorescence 

dye labeled cells (10 μM) in the presence of…..

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