Cell Navigator NBD神经酰胺高尔基体染色试剂盒 绿色荧光
| Ex (nm) | 467 | Em (nm) | 538 |
| 分子量 | - | 溶剂 | - |
| 存储条件 | - |
高尔基体是大多数真核细胞胞质内囊泡和折叠膜的复合体，参与分泌和细胞内运输。它修饰内质网（ER）中内置的蛋白质和脂质，并准备将其输出到细胞外。它还在脂质在整个细胞中的运输和溶酶体的形成中起着重要作用。该Cell Navigator™NBD神经酰胺高尔基染色试剂盒提供了一种简便，快速的方法，可以选择性地对活细胞或醛固定细胞中的高尔基染色。将C6 NBD神经酰胺与牛血清白蛋白（C6-NBD-Ceramide-BSA）形成复合物给予细胞。高尔基体通过形成相应的荧光代谢产物而被染色。
适用仪器
| 荧光显微镜 | |
| Ex: | 488滤波片组 |
| Em： | 525 滤波片组 |
| 推荐孔板: | 黑色透明底板 |
| 滤波片组: | FITC滤波片组 |
样品分析方案
概述
根据需要处理细胞
加入C6 NBD Ceramide工作液，室温或37℃孵育15~30分钟
更换染色缓冲液，室温或37°C孵育15~30分钟
使用 FITC 滤光片组在显微镜下观察
可选：用 4% 甲醛固定细胞
储备溶液配制
除非另有说明，所有未使用的储备溶液应分成一次性等份，并在制备后储存在-20°C。避免反复冻融循环
注意：将未使用的 C6 NBD 神经酰胺储备溶液以一次性等分试样形式储存在 -20o C 下。避免冻融循环。
可选：将 10 µL Hoechst 33342（组分 D）添加到 1 mL NBD 神经酰胺工作溶液中以进行核染色。使用 DAPI 滤光片组在荧光显微镜下观察。
操作步骤
- 根据需要铺板并处理细胞。
- 直接在细胞培养基中添加等体积的 C6 NBD 神经酰胺工作溶液。注意：如果 96 孔板中有细胞，细胞培养基为 100 µL/孔，则添加 100 µL/孔的 C6 NBD 神经酰胺工作溶液。
- 在室温或37°C下孵育15至30分钟。
- 取出 C6 NBD 神经酰胺工作溶液，并用 100 µL/孔的染色缓冲液（组分 B）替换。
- 室温或37℃孵育10~15分钟。
- 使用 FITC 滤光片组在荧光显微镜下观察。
- 可选：除去步骤 4 中的染色缓冲液，并向每个孔中添加 100 µL/孔/96 孔板的 4% 甲醛固定缓冲液（未提供）。
- 注意：对于非贴壁细胞，添加所需量（例如 2X106 个细胞/mL）的 4% 甲醛固定缓冲液。将板在室温下孵育 20 至 30 分钟。除去固定剂。用 PBS 清洗细胞 2-3 次，然后更换为 100 µL/孔的染色缓冲液（组分 B）。
- 注意：Cell Navigator NBD 神经酰胺高尔基体染色试剂盒也适用于醛固定细胞。固定后，按照染色方案步骤 2 至 6 进行操作
图示
图1.HeLa细胞中GGR169神经酰胺高尔基染色的荧光图像。在37°C下，用100μL具有Hoechst 33342的GGR169-神经酰胺工作溶液对细胞染色20分钟。
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