In Vivo Imaging용 Near-IR CF Dye
현존하는 dye에 비해 훨씬 밝은빛을 내는 형광 dye로, 빛에 노출에도 형광이 안정하게 유지되며 in-vivo imaging에도 사용할수 있습니다.
- Brightness - 기존의 Alexa Fluor나 Cy dye에 비해 훨씬 밝은 빛을 냅니다.
- Photostability - 빛에 여러 번 노출되어도 안정하여 photobleaching이 없습니다.
- Specificity and in-vivo Stability - CF로 라벨링을 함으로써 specificity가 더 높고, in vivo imaging시 반감기가 향상됩니다.
- pH Insensitivity - wide pH 범위에서도 높은 형광을 유지합니다.
- Solubility - water나 유기용매에 매우 잘 녹습니다.
- Compatibility - 기존에 사용되는 일반적인 dye와 같은 excitation source, filter를 사용할수 있습니다.
- 편리성 - 항체 labeling, purification, sterile filter까지 모두 제공됩니다.
- 제공량 - 1mg의 항체를 3번 레이블링 할 수 있는 시약이 제공됩니다.
Figure 1: Stability of Cy7, AlexaFluor750 and CF750 dyes. Shown are the absorption spectra of the respective dyes before (black line) and after (gray line) 30 minutes of exposure to sunlight.
Figure 2: Jurkat cells were stained with isotype or mouse anti-human CD3 antibody followed by goat anti-mouse APC-AlexaFluor750 (Invitrogen) or CF750 using the amount of antibody shown. Flourescence was analyzed using a BD LSR Ⅱ flow cytometer equipped with a 633 nm laser and 780/60 nm PMT detector. Bars represent the relative fluorescence values of the geometric means.
Figure 3: Bars represent the signal-to-noise ratio of the relative fluorescence values of the geometric means from Figure 2.
Figure 4: Goat anti-mouse lgG was labeled with CF750, AlexaFluor750 or Cy7, respectively. Fluorescence values were measured on a Hitachi F-4500 fluorometer. The normalized fluorescence values are shown above.
Figure 5: Goat anti-mouse lgG was with CF750, AlexaFluor750 or Cy7. Absorbance of the conjugated dyes were normalized to their respective absorbance max. Cy7 and AlexaFluor750 have large shoulder peaks which are indicative of dye aggregation and to not contribute to overall fluorescence intensity.
Figure 6: Jurkat cells were stained with isotype or mouse anti-human intracellular CD3 antibody followed by 1 ug of goat anti-mouse lgG conjugated with Cy5.5, AlexaFluor680, CF680 or DyLight680. Fluorescence was detected by a BD FACS Calibur in the FL4 channel. The bars represent the signal-to-noise ratio of CD3 positive fluorescence to isotype using similar degrees of labeling (DOL).
Figure 7: HeLa cells were stained with mouse alpha-tubulin antibody followed by CF680 goat anti-mouse lgG. Images were captured using an Olympus mercury arc lamp microscope equipped with a Cy5 filter set, CCD camera and ImagePro Express softwere.
Figure 8: Jurkat were stained with isotype or intracellular CD3 antibody followed by 1ug of goat anti-mouse lgG conjugated with CF770, DyLight 800 or IRDye800CW, respectively. Fluorescence was measured on a BD LSR Ⅱ equipped with a 633 nm laser and 780/60 nm PMT detector. The bars represent the relative flurorescence values of the geometric means.