辫颁补蝉辫补蝉别3-蝉别苍蝉辞谤哺乳黄色荧光信号通路质粒

产物信息
基本信息
质粒介绍
质粒图谱
质粒序列

产物信息

产物货号	产物名称	产物规格	优惠价
XY1712	辫颁补蝉辫补蝉别3-蝉别苍蝉辞谤哺乳黄色荧光信号通路质粒	20μ濒	?请加蚕蚕询价

使用说明

信裕质粒平台的各批次质粒菌株发货前均经过严格的多重验证，如存在质量问题，请在收到产物的三个月内通知我司。收到质粒后请短暂离心，加入20μ濒 ddH2O溶解质粒，取2μl转化至对应感受态中，挑取单克隆重新提取质粒后使用。如需获取其他详细信息请登录我司官网查询。

基本信息

原核抗性:	卡那霉素碍补苍
筛选标记:	遗传霉素骋418
克隆菌株:	大肠杆菌顿贬5α
培养条件:	37℃

质粒介绍

pCaspase3-sensor是一个哺乳细胞黄色荧光蛋白信号通路报告质粒。pCaspase3-Sensor Vector can be used to detect the onset of caspase-3 activity in mammalian cells. This vector encodes the enhanced yellow-green variant (EYFP) of the Aequorea victoria green fluorescent protein (GFP) fused at the 3' end to three copies of the nuclear localization signal (NLS) of the simian virus 40 large T-antigen (1,2). At the 5' end the gene contains a sequence encoding the nuclear export signal (NES) of the Map Kinase Kinase (MAPKK;3). The NES is separated from the EYFP coding region by a 36-nucleotide sequence encoding the region of Poly (ADP-ribose) polymerase (PARP) cleaved by caspase-3. The complete coding sequence for this fusion protein is human codon-optimized (4). Because the NES of MAPKK dominates the NLS, the full-length fluorescent fusion protein distributes to the cytosol. If caspase-3 is activated, the NES will be cleaved from the fusion protein and the truncated EYFP-NLS fusion will translocate to the nucleus via the NLS. The translocation of the fluorescent protein from the cytosol to the nucleus indicates caspase-3 activity at a cellular level. The fluorescence excitation maximum of EYFP is 513 nm; the emission spectrum has a peak at 527 nm (in the yellow-green region). The Em of EYFP at 513 nm is 36,500 cm–1M–1 and its quantum yield is 0.63 (5), resulting in a bright fluorescent signal. The fluorescence intensity is roughly equivalent to that of EGFP.
The vector contains an SV40 origin of replication and a neomycin resistance (Neor) gene for selection (using G418) in eukaryotic cells. A bacterial promoter (P) upstream of Neor expresses kanamycin resistance in E. coli. The vector backbone also provides a pUC19 origin of replication for propagation in E. coli and an f1 origin for single-stranded DNA production.
The pCaspase3-Sensor Vector can be introduced into mammalian cells using any standard transfection method. If desired, stable transfectants can be selected using G418 (6).pCaspase3-Sensor can be used to detect the onset of caspase-3 activity in mammalian cells. This vector allows visual monitoring of caspase-3 activity in the cell via fluorescence microscopy. If the fluorescent fusion protein encoded by the pCaspase3-Sensor vector is in the cytosol, caspase-3 is not active. However, if caspase-3 becomes active, the dominant nuclear export signal (NES) is cleaved from the fusion protein and the truncated fluorescent protein will become localized to the nucleus via the nuclear localization signal.

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