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产品资料 PD-40704ES

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现货亚细胞研究翌圣生物 Yeasen

Fluo-4,AM钙离子荧光探针(细胞可渗透) Fluo-4钙离子探针

Fluo-4, AM, Cell Permeant 细胞膜可渗透钙离子荧光探针

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荧光检测钙离子信号通路研究药物筛选与评价

产品编号40704ES 可选规格3 种 来源翌圣生物 · 官网

产品编号 40704ES50

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数量 1

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货号规格价格状态

40704ES50 50 μg $268.00促~~$276.00~~ 现货询价

40704ES72 5× 50 μg $928.00促~~$976.00~~ 现货询价

40704ES80 1 mg $2,448.00促~~$2,576.00~~ 现货询价

提交询价价格为协议价，提交后由业务确认；大包装可申请试用装。

产品概述

技术参数

品牌翌圣生物 Yeasen

货号 / 型号40704ES

规格50 μg

产地翌圣生物科技(上海)股份有限公司

技术平台 荧光检测

应用方法 钙离子信号通路研究用于细胞钙信号转导机制研究 药物筛选与评价评价化合物对细胞内钙离子浓度的影响

检测方法 荧光探针法

生物特性参数 探针类型细胞膜可渗透钙离子荧光探针 作用机制AM酯穿透细胞膜后被酯酶剪切滞留胞内，结合Ca2+产生强荧光 靶标通路钙离子信号通路 荧光增强倍数结合钙离子后荧光增强60-80倍 细胞膜通透性可渗透细胞膜（AM酯形式）

样本类型细胞

技术参数 CAS号273221-67-3 分子式C51H50F2N2O23 分子量1096.94 纯度≥90% (HPLC) 外观橙红色粉末 Ex/Em494/516 nm (结合Ca2+) 储存条件-20℃避光干燥保存，可保存1年 运输条件室温运输

实验类型 细胞内钙离子浓度检测检测细胞内钙离子浓度动态变化 荧光显微成像对细胞内钙信号进行荧光成像观察

常见问题

Q：有哪些比较常见的可见光激发Ca2+荧光探针？

A：Fluo-3 是最常用的可见光激发 Ca2+荧光探针、Fluo-4 是钙指示剂Fluo-3 的升级版本、Rhod-2 是另一种可见光激发的高亲和力Ca2+指示剂。

Q：几种探针的应用的位置？

A：如果是测定胞质内的钙离子建议选择 Fluo-3、4，如果是测定细胞器内较浓钙离子的实验建议采用Rhod-2

Q：通过流式细胞仪可以使用什么指示剂染料来测量钙通量？

A：Indo-1 是流式细胞术的首选染色剂，使用单一激光器进行激发（通常是氩离子激光器的 351-364 nm 谱线）并监测两种发射，Indo-1 的发射最大值从无 Ca2 +培养基中〜475nm 转变为当染料被 Ca2+结合饱和时的〜400nm。Indo-1 比较适用于多色荧光应用。

Q：荧光探针用什么缓冲液稀释？

A：工作液用最好是使用无血清培养基、HBSS 或是 PBS 等缓冲液进行稀释，稀释后的探针工作液即

可直接加入细胞进行检测。

Q：荧光探针装载完多久后观察？

A：为减少各种可能的误差，尽量缩短探针装载后到仪器测定所用的时间(刺激时间除外)。

Q：Fura-2 为什么需要两次不同的激发和发射测定？

A：结合 Ca2+后的最大激发波长从结合前的 380nm 向 340nm（Ca2+饱和时）偏移，其发射荧光强度与结合 Ca2+的浓度存在定量关系。一般用 340nm 和 380nm 波长激发 Fura-2，在 510nm 处检测发射波长。通过使用与两种激发对应的荧光强度比率来计算细胞内的钙离子浓度，这种比率测量方法可以消除不同细胞样品间荧光探针装载效率的差异，荧光探针的渗漏，细胞厚度差异等一些误差因素。

Q：我需要用 Fluo-4 标记细胞进行钙通量分析。标记后多久可以保留染料？

A：将染料加入细胞后，细胞内酯酶从染料中除去'AM'部分。当'AM'基团被移除时，染料能够结合钙和发出荧光。由于染料不与任何细胞成分共价结合，因此可能会主动从细胞中流出。流出速度取决于细胞的固有特性，培养条件和其他因素。染料可能会保留数小时，数天甚至数周，或在几分钟内丢失。建议染色后应立即进行检测，也可以使用丙磺舒限制主动外流造成的损失。

Q：加载 Fluo-4 染料检测钙后可以固定细胞吗？

A：不能。因为 Fluo-4 不与任何细胞成分共价结合并且固定会损害膜，染料将不会被细胞保留。

Q：对于人肝星状细胞LX-2建议的Fluo-4工作浓度和孵育时间有吗？

A：参考文献PMID:27473374，可使用0.5uM Fluo-4，AM于37℃孵育15min。

文档与文献

已发表文献

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