研选Cell Cycle Staining Kit 细胞周期检测 试剂盒 - 联科生物 MultiSciences 百篇参考文献引用 流式细胞周期检测步骤
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500篇文献引用,你还担心什么呢?独创活细胞一步法检测:收集 - 洗涤 - 染色,简单快速,更小CV
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文献和实验
研选推荐
库存 :500
英文名 :Cell Cycle Staining Kit
CAS号 :无
保质期 :1年
供应商 :联科生物
保存条件 :2 - 8℃ 保存
规格 :50T
联科生物有两种细胞周期检测试剂盒,在组分上,CCS01是DNA染色液和破膜剂的混合装;CCS012 为DNA染色液和破膜剂的独立包装。在用途上,CCS01可用于一步法活细胞细胞周期检测,固定细胞也 可用,但推荐用CCS012效果更佳;CCS012可用于活细胞和固定细胞的细胞周期检测。
产品图片
产品简介
碘化丙啶(Propidium iodide, PI)可插入碱基之间而与DNA结合,其对碱基几乎没有序列偏好,每4 - 5个碱基对可结合一个染料分子。PI也能与RNA结合,需要用核酸酶处理来区分RNA和DNA染色。一旦染料与核酸结合,其荧光会增强20 - 30倍。虽然其摩尔吸收率(吸光系数)相对较低,PI仍然具有较大的stokes位移,从而在滤光片选择恰当的情况下,仍能同时检测细胞核DNA和荧光标记的抗体。PI可用于荧光显微镜、激光共聚焦、流式细胞仪和荧光计。
PI为膜不通透性,被活细胞排斥,可用于鉴定死细胞和多色荧光复染。下面提供的操作步骤与很多细胞标记技术兼容,如直接或间接抗体检测技术、mRNA原位杂交、特异性的细胞结构染料等。在组织染色中可能要作修改。
本试剂盒包含50 ml DNA染色液和500 μl破膜剂,足够用于50次检测。
产品特点
- 即用型,操作简单
- 活细胞/固定细胞检测
- 常备现货
使用方法
一、可当天检测的活细胞样本
1. 收集2 × 105 - 1 × 106个细胞,离心弃上清。用PBS洗涤一次,离心弃上清。
2. 加入1 ml DNA Staining solution和10 μl Permeabilization solution,涡旋振荡5 - 10秒混匀。室温避光孵育 30分钟。
3. 选择最低上样速度,在流式细胞仪上进行检测。如需在荧光显微镜下观察,则离心细胞,弃上清,加 入新鲜缓冲液重悬。滴加1滴细胞悬液至载玻片,盖上盖玻片后使用合适的滤光片进行观察。
二、当天无法检测的活细胞样本
1. 按照下述方法固定,或其他合适的方法固定
a. 收集2 × 105 - 1 × 106个细胞,离心弃上清。轻弹管壁,使沉淀重悬在残余的液体中,加入1 ml 室温 下的PBS。
b. 将细胞缓慢加入至3 ml无水乙醇(-20℃预冷)中,边加边高速搅拌。-20℃固定过夜,可保存数月。
2. 检测当天,将固定细胞离心,弃去乙醇,轻弹管壁使沉淀松散,加入2 - 5 ml 室温下的PBS,放置15 分钟使细胞再次水化。离心,弃上清。
3. 加入1 ml DNA staining solution,涡旋振荡5 - 10秒混匀。室温避光孵育30分钟。
4. 选择最低上样速度,在流式细胞仪上进行检测。如需在荧光显微镜下观察,则离心细胞,弃上清,加 入新鲜缓冲液重悬。滴加1滴细胞悬液至载玻片,盖上盖玻片后使用合适的滤光片进行观察。
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