lucernatechnologies：OBI

3,5-difluoro-4-hydroxybenzylidene-imidazolinone-2-oxime-1-benzoimidazole (OBI)

分子式:C19H13F2N5O3

分子量:397.34

CAS NAME/NUMBER:

Fluorescence Spectra

Absorption and fluorescence emission spectra of RedBroccoliTM/OBI in pH 7.4 buffer.

PRESENTATION: 冻干染料

质量保证：产品由以下方式分析1H NMR 和 LC-MS，并通过 HPLC 以 >95% 的纯度提供。

使用声明：本产品仅供研究使用，不得用于任何其他目的，包括但不限于未经授权的商业用途、体外诊断用途、离体或体内治疗用途或任何类型的消费或对人类或动物的应用。由于荧光团和适配体的高度特异性，我们无法预测或对 Lucerna 如何负责TM的产品将在其客户的系统中运行。使用 Lucerna 的研究人员TM的产品应进行优化研究，以达到其预期应用的最佳结果。

数据

Figure 1. Red BroccoliTM-OBI complex exhibits increased photostability in cells and in vitro. In-cell photostability of Red BroccoliTM-fluorophores (OBI versus BI, 20 µM each) was assessed by continuous illumination of Red BroccoliTM-expressing cells. Red BroccoliTM-OBI maintained >75% of its red fluorescence for over 10 s, while Red BroccoliTM-BI lost ~50% of its green fluorescence within ~0.2 s. Scale bar, 10 µm.

Figure 2. The enhanced cellular stability of OBI enables imaging of endogenous SAM dynamics in mammalian cells using a Red BroccoliTM-based SAM biosensor. (a) A schematic diagram of the SAM sensor used in this experiment. (b) HEK293T cells expressing either the Red BroccoliTM-based SAM sensor (top) or Red BroccoliTM alone (bottom) were imaged with 10 µM OBI. Sensor-expressing cells treated with 50 mM cycloleucine at 0 min to block SAM biosynthesis exhibited rapidly decreases in cellular fluorescence. After 120 min, the media was replaced with cycloleucine-free media (“Wash out") and fluorescence recovered to baseline levels over time. Cells expressing only Red BroccoliTM maintained consistent red fluorescence over time. Scale bar, 20 µm.

Figure 3.  BI and OBI are “plug-and-play" fluorophores for BroccoliTM and Red BroccoliTM. HEK293T cells expressing Red Broccoli, Broccoli, or control RNA were incubated with 10 µM OBI (a-c) or BI (d-f). OBI exhibited red fluorescence in  both Red Broccoli- and Broccoli-expressing cells and BI exhibited green fluorescence in both Red Broccoli- and Broccoli-expressing cells. But no fluorescence was seen in control RNA-expressing cells with either fluorophore. Cell nucleus was labeled with Hoechst-33342 stain (blue) and imaged with DAPI and a FITC filter cube for BI or TRITC filter cube for OBI. Scale bar, 10 µm.

Reference

Imaging intracellular S-adenosyl methionine dynamics in living mammalian cells with a genetically encoded red fluorescent RNA-based sensors. Li X, Mo L, Litke JL, Dey SK, Suter SR, Jaffrey SR. J Am Chem Soc. 2020 Aug 19;142(33): 14117-14124.