保存条件 RNase A Reagent 适当分装后-20°C 可保存 1 年。Cycle Blue Reagent -20°C 避光可保存 1 年。 检测原理 Elabscience®的 Cell Cycle Assay Kit (Blue Fluorescence)是通过检测 DNA 含量来检测细胞周期的试 剂盒。本试剂盒可应用于培养细胞(悬浮、贴壁)的 DNA 含量(细胞周期)检测。 细胞周期(cell cycle)是指连续分裂细胞从一次有丝分裂结束到下一次有丝分裂结束所经历的整个 过程。在这个过程中,细胞遗传物质复制并加倍,且在分裂结束时平均分配到两个子细胞中去。细胞周 期可以分为分裂间期(interphase)和有丝分裂期(M phase),分裂间期又可以细分为休眠期(G0),DNA 合成前期(G1),DNA 合成期(S),DNA 合成后期(G2)。利用细胞内 DNA 能够和某些特定荧光染 料(如 Cycle Blue)结合的特性,细胞各个时期其 DNA 含量不同从而结合的荧光染料的量不同,通过 流式细胞仪检测荧光强度即可反映细胞周期变化情况。 Cycle Blue 染色后,假设 G0/G1 期细胞的荧光强度为 1,那么含有双份基因组 DNA 的 G2/M 期细 胞的荧光强度的理论值为 2,正在进行 DNA 复制的 S 期细胞的荧光强度为 1~2 之间。凋亡细胞由于细 胞核发生浓缩以及 DNA 片段化(DNA fragmentation)导致部分基因组 DNA 片段在染色过程中丢失,因 此凋亡细胞在 Cycle Blue 染色后呈现明显的弱染,即荧光强度小于 1,在流式检测的结果图上出现所谓 的 sub-G1 峰,即凋亡细胞峰。 Detection principle Elabscience's® Cell Cycle Assay Kit (Blue Fluorescence) is a test kit that measures the cell cycle by measuring DNA content. This kit can be applied to the detection of DNA content (cell cycle) of cultured cells (suspension, adherent). The cell cycle refers to the process that a cell undergoes from the end of one mitosis to the end of the next mitosis when it divides continuously. In this process, the genetic material of the cell is duplicated and doubled, and at the end of the division it is equally distributed to the two daughter cells. The cell cycle can be divided into interphase and mitosis, which can be subdivided into dormant phase (G0), prophase of DNA synthesis (G1), phase of DNA synthesis (S), and phase of DNA synthesis (G2). Taking advantage of the fact that intracellular DNA can bind to certain fluorescent dyes (such as Cycle Blue), the amount of fluorescent dyes bound to cells varies depending on the amount of DNA content at different stages, and the fluorescence intensity can be detected by flow cytometry to reflect the changes in the cell cycle. After Cycle Blue staining, assuming that the fluorescence intensity of cells in the G0/G1 phase is 1, then the theoretical value of the fluorescence intensity of G2/M phase cells containing double genomic DNA is 2, and the fluorescence intensity of cells in S phase undergoing DNA replication is between 1~2. Apoptotic cells are lost in the staining process due to the condensation of nuclei and DNA fragmentation, so the apoptotic cells show obvious weak staining after Cycle Blue staining, that is, the fluorescence intensity is less than 1, and the so-called sub-G1 peak, that is, the apoptotic cell peak, appears on the result map of flow cytometry.