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: 凍干染料

質(zhì)量保證：產(chǎn)品由以下方式分析1H NMR 和 LC-MS，并通過 HPLC 以 >95% 的純度提供。

使用聲明：本產(chǎn)品僅供研究使用，不得用于任何其他目的，包括但不限于未經(jīng)授權(quán)的商業(yè)用途、體外診斷用途、離體或體內(nèi)治療用途或任何類型的消費或?qū)θ祟惢騽游锏膽?yīng)用。由于熒光團和適配體的高度特異性，我們無法預(yù)測或?qū)?Lucerna 如何負責(zé)TM的產(chǎn)品將在其客戶的系統(tǒng)中運行。使用 Lucerna 的研究人員TM的產(chǎn)品應(yīng)進行優(yōu)化研究，以達到其預(yù)期應(yīng)用的最佳結(jié)果。

數(shù)據(jù)

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.