Click-iT™ Plus EdU Alexa Fluor™ 488 Imaging Kit
货号:C10637
规格:1 kit
价格:8225
产品类型:细胞增殖活性检测
品牌:Thermo Fisher
物种:恒河猴/小鼠/大鼠
宿主:小鼠
抗体亚型:IgG
荧光染料:Alexa Fluor 488
Click-iT Plus EdU 细胞增殖成像试剂盒已优化并应用于荧光显微镜,且这是一种优于传统增殖检测的方法。本测定试剂盒中经修饰的胸腺嘧啶核苷类似物 EdU(5-乙炔基-2'-脱氧尿苷,一种胸腺嘧啶核苷的核苷类似物)能有效的掺入新合成的 DNA 并且通过明亮的、非感光的 Alexa Fluor™ 染料以快速、高特异性、温和的点击反应进行荧光素标记。• 简单 — 与传统方法相比,第一次试验及每次试验花费的时间更少• 高效 — 无需变性步骤或严格的处理• 结果内容丰富 — 改善了细胞形态、抗原结构、GFP 荧光信号和 DNA 完整性的保存• 一致性 — 检测不依赖不同抗体批次该试剂盒包含标记和检测掺入的 EdU 及对贴壁细胞样品进行细胞周期分析所需的所有成分。用于细胞周期分析时,试剂盒包含蓝色荧光 Hoechst 33342 染料。本款试剂盒包含的试剂足够供每次检测使用 500 µL 的反应缓冲液标记50个 18x18 盖玻片。避免与 BrdU 方法相关的严格处理测量细胞增殖变化是评估细胞健康、确定基因毒性和评估抗癌药物的基本方法。较准确的方法是直接检测 DNA 的合成。传统的方法利用核苷类似物 BrdU(5-溴-2'-脱氧尿苷,一种胸腺嘧啶核苷的核苷类似物)掺入新转录的 DNA。样品掺入后用严格的方法处理(HCl、加热或酶)使 DNA 变性并将 BrdU 分子暴露于抗 BrdU 抗体进行检测。然而,使用 BrdU 方法检测细胞增殖较为耗时,且难以一致地进行。这种方法所需的严格处理会对样品完整性、细胞形态、图像质量和多通路能力产生不利影响。Click-iT™ Plus EdU 检测是基于核苷类似物 EdU 掺入 DNA 以测定新 DNA 合成率。检测是通过催化的“点击”反应完成的,该反应通常在30分钟内完成。点击反应使用生物正交(生物特有)部分以荧光标记增殖的细胞,产生较低的背景和较高的检测灵敏度。由于温和的反应条件,Click-iT™ Plus 检测可精确的确定细胞增殖,同时保留细胞形态、DNA 完整性、抗原结合位点和 GFP 荧光信号。DNA 完整性的保留可使 DNA 着色,包括用于细胞周期分析的染色。 |
数据: |
图1.Signal from RFP protected and proliferating cells detected using the Click-iT™ Plus EdU Alexa Fluor® 488 Imaging KitHeLa cells were transduced with CellLight® Early Endosome RFP (C10587). After an overnight incubation, proliferation was detected using the Click-iT™ Plus EdU Alexa Fluor® 488 Imaging Kit (C10637). Red fluorescent signal is from the transduced RFP construct and the green signal represents proliferating cells.图2.mCherry signal preserved in multipotent otic progenitor cells labeled with EdUGenetically modified induced multipotent otic progenitor cells expressing mCherry were initially cultured in DMEM/F12, B27 containing 20 ng/ml of bFGF. To promote neuronal differentiation, cells were plated onto 1.5 glass coverslips coated with 10 µg/mL poly-D-lysine and 10 µg/mL laminin and cultured in the same media for 7 days without bFGF. To incorporate the nucleoside analog EdU and identify proliferating cells, cultures were incubated with 1 µM EdU for 2 hrs before fixation. Media was removed from the cultures and cells were fixed in 1X PBS containing 4% formaldehyde for 15 minutes at RT. Cells were washed in 1X PBS containing 3% BSA and the cell membrane permeabalized in 1X PBS containing 0.5% Triton X-100 for 20 minutes at RT. To label the EdU, cells were washed with 1X. The top and bottom rows represent use of the Click-iT® Plus EdU Alexa Fluor® 488 Imaging Kit and the Click-iT® EdU Alexa Fluor® 488 Imaging Kit, respectively. |
相关产品: |
▪细胞周期检测▪细胞凋亡检测▪细胞增殖活性检测 |
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