DiSBAC2(3)细胞膜电位荧光探针

DiSBAC2(3)细胞膜电位荧光探针；DiSBAC2(3); [Bis-(1,3-diethylthiobarbituric acid)trimethine oxonol]
DiSBAC2(3)是一种带有长波长激发和发射光灵敏的膜电位探针。它是一种慢响应的探针，用于测定细胞跨膜电位。总的来说，慢响应探针表现的电位改变依赖于它们跨膜的分布，同时还伴随着荧光的改变。它们巨大的光学反应比快响应的探针要大的多（一般每mV引起1%荧光改变）。慢响应探针，包括羰花青阳离子，罗丹明和氧离子，它们都适合用于细胞非应激性平均膜电位的检测；这些非应激性细胞活动通常由细胞呼吸作用，离子通道渗透性，药物结合和其它一些因素导致。

规格	25 mg	产品形式	粉末
Ex (nm)	535	Em (nm)	560
分子量	436.55	溶剂	DMSO

储存条件：-20℃
操作说明（仅供参考）
1.Prepare cells:
1.1 For adherent cells: Plate cells overnight in growth medium at 40,000 to 80,000 cells/well/100 µL for 96-well plates or 10,000 to 20,000 cells/well/25 µL for 384-well plates.
1.2 For non-adherent cells: Centrifuge the cells from the culture medium and then suspend the cell pellets in equal amount of HHBS and MP dye-loading solution (see Step 2.2 below) at 125,000 to 250,000 cells/well/100 µL for 96-well poly-D lysine plates or 30,000 to 60,000 cells/well/25 µL for 384-well poly-D lysine plates. Centrifuge the plates at 800 rpm for 2 minutes with brake off before the experiments.
Note: Each cell line should be evaluated on an individual basis to determine the optimal cell density for the intracellular calcium mobilization.
2.Prepare DiSBAC2(3) dye-loading solution (for 1 plate):
2.1 Prepare a 10 to 30 mM stock solution of DiSBAC2(3) in high-quality, anhydrous DMSO. The stock solution should be used promptly; any remaining solution need be aliquoted and frozen at < -20 oC.
Note: Avoid repeated freeze-thaw cycles, and protect from light.
2.2 Prepare a 2X DiSBAC2(3) dye-loading solution: On the day of the experiment, either dissolve DiSBAC2(3) solid in DMSO or thaw an aliquot of the DiSBAC2(3) stock solution to room temperature. Prepare a 2X working solution of 20 to 40 µM in Hanks and 20 mM Hepes buffer (HHBS) or buffer of your choice, pH 7 with 0.04% to 0.08% Pluronic® F-127 (Cat. # 20053) and 2 mM Trypan Red Plus™ (Cat. # 2456). Mix them well by votexing. This working solution is stable for at least 2 hours at room temperature.
3.Run Membrane Potential Assay:
3.1 Add 100 µL/well (96-well plate) or 25 µL/well (384-well plate) DiSBAC2(3) dye-loading solution (from Step 2.2) into the cell plate.
Note1: If your screen compounds interfere with growth medium and serum factors, replace the growth medium with equal volume of HHBS buffer before adding the DiSBAC2(3) dye-loading solution. Alternatively, cells can be grown in serum-free conditions.
Note 2: Do NOT wash the cells after dye loading.
3.2 Incubate the dye-loading plate in a cell incubator for 30 to 60 minutes.
Note: In some cases, incubation at room temperature for 30 to 60 min may work better.
3.3 Prepare the compound plates by using HHBS or your desired buffer.
3.4 Run the membrane potential assay by monitoring the fluorescence intensity at Ex/Em = 540/590 nm (Cat. # 21414).
Note: It is important to run the signal test before your experiment. Different instruments have their own intensity range. Adjust the signal test intensity to the level of 10% to 15% of the maximum instrument intensity counts. For example, the maximum fluorescence intensity count for FLIPR-384 is 65,000, so the instrument settings should be adjusted to have its signal test intensity around 7,000 to 10,000
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