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TRFS-green
CAS No. : 1513848-14-0
MCE 国际站：TRFS-green
产品活性：TRFS-green 是一种高选择性的荧光探针，用于成像活细胞中的硒蛋白硫氧还蛋白还原酶 (TrxR)。TRFS-green 的最大吸光度在 373 nm 左右。经 TrxR 活化后，最大吸光度约为 440 nm。
研究领域：Metabolic Enzyme/Protease
作用靶点：TrxR
In Vitro: TRFS-green displays a green fluorescence off−on change induced by TrxR-mediated disulfide cleavage and subsequent intramolecular cyclization to liberate the masked naphthalimide fluorophore.
TRFS-green gives a signal at ~480 nm when excited at 377 nm. Pretreatment of living cells with TrxR inhibitors in a dose-dependent manner can inhibit the fluorescence signal of TRFS-green in living cells, thereby facilitating the development of living cell-based screening assays for TrxR inhibitors.
TRFS-green shows no response to GSH or Cys, the predominant thiols in cells, or the endogenous reducing agents, such as vitamin C and NADPH.
Measure TrxR activity in crude cell extracts: Incubate cell lysates with TRFS-green for an appropriate time (usually 1-2 hours), and measure fluorescence intensity by fluorimeter (Em= 540 nm, Ex=440 nm). It is also possible to monitor the time-dependent increase in fluorescence (Em=540 nm, Ex=440 nm) over a given period of time.
Cells treated with TRFS-green (10 μM; 2-4 h) can be observed through the green fluorescent channel of the microscope.
Note: In order to obtain a clear fluorescence image, it is recommended to wash the cells with PBS or fresh medium immediately before taking pictures to remove residual TRFS-green.
TRFS-green is the first fluorescent probe of TrxR with a green emission. However, the response rate of TRFS-green to the enzyme is slow, and it takes more than 2 h to reach the maximal fluorescence signal even using tris(2-carboxyethyl)phosphine (TCEP) as a reducing agent.
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细胞，利用药物 aphidicolin 诱导 293T 为非增殖细胞。实验结果如下： Day3 Day8 Day14 数据显示，IDLV 感染非增殖细胞 14 天依然维持稳定表达能力；而感染增殖细胞时，随着细胞的复制以及传代，8-14 日病毒颗粒已丢失殆尽。 2. 非整合特性验证 分别采用 LV、IDLV 感染 293T 细胞，观察感染后 3、6、8 天的病毒基因组相对含量数值的变化（QPCR 检测病毒基因组与内参 actin 的相对值）： QPCR 检测病毒基因组数据 数据表明，在增殖的 293
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