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Fluorescent Dyes


Fluorescent Dye Overview


Fluorescence is the property of certain substances to emit light. Emission occurs only after absorbing light or electromagnetic radiation at a specific wavelength. Emission light is usually of longer wavelength than the absorbed light. One of the most well-known natural fluorescent probes is Green Fluorescent Protein (GFP) from jellyfish. However, a fluorophore may also be a fluorescent organic compound usually with a low molecular weight (0.2-1 kDa) and typically containing numerous aromatic groups, or plane or cyclic moieties with π bonds. Fluorophores, i.e. either fluorescent proteins or organic dyes, can be directly conjugated to antibodies making them ideal for use in various immunoassays.



 Featured Fluorescent Dye Conjugated Antibodies

 

 

 

 

Fluorescent labeled antibodies can be used to detect target proteins in fluorescent western blots, or dot blots, in FLISA assays, as well as immunofluorescent microscopy, flow cytometry and immunohistochemistry. By using the right combination of fluorescent labeled antibodies multiple targets can be visualized simultaneously in a single assay (fluorescent multiplexing).

 

 

Visible Spectrum

 

 

Primary, or direct, immunofluorescence uses a single antibody conjugated directly to a fluorescent dye. Secondary, or indirect, immunofluorescence uses two antibodies; a primary antibody which recognizes the target biomolecule and binds to it and a secondary antibody conjugated to a fluorescent dye, which recognizes and binds to the primary antibody and indirectly localizes the target for detection. Fluorescent dyes are optimized for excitation and detection in the visible light to near infrared spectrum, 400 nm - 800 nm. The choice of fluorochrome to be used is influenced both by the application and the excitation wavelengths available. The table below provides an overview of fluorescent dies used at Rockland. 

 

Rockland Fluorescent Dye Conjugates

 

 

Fluorescent Label

Color 

Abs (nm) 

Em (nm) 

 

MW (daltons) 

         

DyLight™ 405

Violet

400

420

793

         

Aminomethylcoumarin (AMCA)

Violet Blue

353

442

410

         

ATTO 425

Blue

436

484

498

         

Cy2™

Blue Green

489

505

897

         

DyLight™ 488

Blue Green

493

518

1,011

         

ATTO 488

Green

501

523

981

         

Fluorescein (Fluorescein)

Green

495

528

 390

         

ATTO 532

  Yellow Green

532

553

1,081

         

Cy3™

 Yellow Green

552

565

949

         

DyLight™ 549

 Yellow Green

550

568

982

         

Rhodamine (TRITC)

 Orange

550

570

444

         

R-Phycoerythrin (RPE)

Orange

488

575

240,000

         

ATTO 550

Orange

554

  

576

791

         
Cy3.5™   

Orange Red

581

596

1,286

         

Texas Red®

Red

 596

620

625

         

ATTO 594

Red

 601

627

 1,389

         

Allophycocyanin

Far-Red

650

660

100,000

         

Cy5™

Far-Red

650

667

 975

         

ATTO 647N

Far-Red

 644

669

843

         

DyLight™ 649

Far-Red

646

674

843

         

ATTO 655

 Far-Red

 663

684

887

         

Cy5.5™

Near Infra-Red

678    

703

1,312

         

DyLight™ 680

Near Infra-Red

682

715

950

         

DyLight™ 800

Infra-Red

770

794

1,050

         

 

 

 

 

View All IR Dyes   View all Chemiluminescent Reporters

 

 

 

Rockland conjugates a broad group of secondary antibodies to many of the classic fluorescent markers including fluorescein, rhodamine, Texas Red, CyDyes™ and Phycoerythrin (RPE). All of the conjugates are ideal for various immunofluorescence based assays including fluorescent western blotting, immunofluorescence microscopy, FLISA, and more. Rockland also produces many next generation fluorochrome dyes designed for detection of primary antibodies in multiplex, multi-color analysis. Next generation fluorochrome conjugates (Atto-tec dyes, DyLight™ dyes) offer superior absorption (high extinction coefficient), high fluorescence quantum yield, and superior high photostability.

 

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