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DyLight 680 Antibody Conjugates

Increase multiplexing capacity and reduce background fluorescence with the far-red emission of DyLight 680. The high photostability of DyLight 680 and its excellent compatibility with infrared imaging systems makes it versatile across many use cases.

Western blot analysis of Jurkat cell lysates (#9194) treated with either U0126 (MEK 1/2 inhibitor) #9903 or TPA (12-O-Tetradecanoylphorbol-13-Acetate) #4174, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/204) (D13.14.4E) XP® Rabbit mAb #4370 detected with Anti-rabbit IgG (H+L) (DyLight 800 Conjugate) #5151 (green) and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107 detected with Anti-mouse IgG (H+L) (DyLight 680 Conjugate) (red).

Western blot analysis of Jurkat cell lysates #9194 treated with either U0126 (MEK 1/2 inhibitor) #9903 or TPA #4174, using Phospho-p44/42 MAPK (Erk1/2) (Thr202/204) (D13.14.4E) XP® Rabbit mAb #4370 detected with Anti-rabbit IgG (H+L) (DyLight 800 Conjugate) #5151 (green) and p44/42 MAPK (Erk1/2) (3A7) Mouse mAb #9107 detected with Anti-mouse IgG (H+L) (DyLight 680 Conjugate) #5470 (red).

Attributes

Excitation: 692 nm

Emission: 712 nm

Laser Line: 628-640 nm

Common Filter Set: 695/40

Brightness: 4

DyLight 680 Excitation Spectrum

The dashed line shows the excitation spectrum of DyLight 680, while the solid line shows its emission spectrum. When performing multiplexing experiments, it is best to select fluorophores with emission spectra that do not overlap. DyLight 680 has a broad excitation profile, allowing it to be excited by a range of NIR light sources.



Applications in Research

Researchers benefit from several key advantages of DyLight 680:

  • Being a far-red dye with negligible channel bleed-through, DyLight 680 avoids emission overlap with red, yellow, green, and blue dyes, making it a good choice for multiplexing experiments.
  • By using DyLight 680 with a second near-infrared fluorophore, such as DyLight 800, researchers can perform dual-color Western blotting, allowing for the simultaneous detection and quantification of multiple proteins on the same membrane.
  • DyLight 680’s consistent brightness and photostability allow for accurate quantitative imaging, making it useful for applications requiring precise measurements, such as protein quantification.

The superior photostability of DyLight 680 enables several applications, including:

In-Cell Western™ analysis of A549 cells exposed to varying concentrations of U0126 (MEK1/2 Inhibitor) #9903 for 3 hours, followed by TPA (Phorbol-12-Myristate-13-Acetate) #9905 stimulation for 30 minutes. With increasing concentrations of U0126, a significant decrease (~5 fold) in Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 signal as compared to the TPA-stimulated control was observed. When using phospho-Erk as a measurement, the IC50 of this compound was 1.9 μM. Data and images were generated on the LI-COR Biosciences Odyssey Infrared Imaging System using Anti-rabbit IgG (H+L) (DyLight 680 Conjugate).

Accurately quantify low-expressed proteins

In-Cell Western analysis of A549 cells exposed to varying concentrations of U0126 (MEK1/2 Inhibitor) #9903 for 3 hours, followed by TPA #9905 stimulation for 30 minutes. With increasing concentrations of U0126, a significant decrease (~5 fold) in Phospho-p44/42 MAPK (Erk1/2) (Thr202/Tyr204) (D13.14.4E) XP® Rabbit mAb #4370 signal as compared to the TPA-stimulated control was observed. Data and images were generated using Anti-rabbit IgG (H+L) (DyLight 680 Conjugate) #5366.

Commitment to Quality

Get accurate and reliable experimental results with functionally validated, ready-to-use DyLight 680 antibody conjugates from CST. Stringent and reproducible manufacturing process results in high-quality antibody conjugates produced to the same standards you've trusted for over 25 years. For custom or made-to-order conjugates, we rely on our decades of experience, thorough biophysical characterization, and parent antibody validation to deliver a product with high specificity and sensitivity. Every order comes with comprehensive product documentation and access to expert technical support.

Related or Alternative Fluorophores

Cy®5.5

Alexa Fluor® 680

IRDye 680

For Research Use Only. Not for Use in Diagnostic Procedures.

Cell Signaling Technology, CST, and XP are trademarks or registered trademarks of Cell Signaling Technology, Inc. Cy is a registered trademark of GE Healthcare. Alexa Fluor is a registered trademark of Life Technologies Corporation. All other trademarks are the property of their respective owners. Visit cellsignal.com/trademarks for more information.