BIO AND SMALLER MOLECULES

  •    
  • Home
  • CHROMIS FLUORESCENT DYES

CHROMIS FLUORESCENT DYES

Introduction

CHROMIS is a product line of fluorescent dyes for the conjugation of bio and smaller molecules.
The line covers the full range from 400 to 800 nm.
The dyes are available with different properties and reactivities.
After the bio-conjugation, the CHROMIS dyes can be used in all the most common applications for molecular biology, including Western blotting, Flow Cytometry and Fluorescent Microscopy.

Features

  • Ready to use for conjugation
  • High brightness
  • High purity
  • Availability of different properties and reactivities
  • Customized derivatization, including conjugation with bio and smaller molecules

The names of the fluorescent dyes in the CHROMIS series are formulated using different abbreviations:

  • CHROMIS: Name of the class of fluorophores developed by Cyanagen.
  • Number: Three-digit number indicates the approximate absorption peak wavelength in nanometers.
  • Charge: A single capital letter indicates the overall charge state of the molecule (C as cationic, A as anionic, Z as zwitterionic and N as neutral).
  • Reactive moiety: A short description of the reactive moiety at the end of the spacer arm.

(*) = Products available only on request

CHROMIS

Ex-Max (nm)

Excitation
laser line

Commonly used filter sets

Em-Max (nm)

Molar
absorbance
(M-1cm-1)

Quantum yield

Quencher

Class

Spectrally
comparable
dyes

425 N

425

Diode 405 nm blue-violet

DAPI

476

37,000

Not available

QS35

COUM

PromoFluor 415

500 N

496

Argon 488

nm blue-green

Diode 473 nm blue

FITC

506

88,000

1.00

QSY7

BDP

Alexa Fluor 488
ATTO 488
BODIPY FL
Cy2
Fluorescein (FITC)
Promo Fluor 488/505

515 N (*)

515

Argon 488 nm blue-green

Diode 473 nm blue

R6G

539

50,000

0.80

QSY7

BDP

Alexa Fluor 514
6-JOE/6-TET
Promo Fluor 520

530 N

529

Nd YAG

Diode 532 nm green

R6G

561

54,000

0.70

QSY9

BDP

BODIPY R6G
Alexa Fluor 532
Tetramethylrhodamine (TMR)

550 C (*)

548

Nd YAG

Diode 532 nm green

He-Ne 543 nm green

TRITC

567

150,000

0.04

C5Q

CYAN

Alexa Fluor 546/555
BODIPY TMR
Cy3
DyLight 549
Promo Fluor 546/555
Tetramethylrhodamine (TMR)

550 Z (*)

550

Nd YAG

Diode 532 nm green

He-Ne 543 nm green

TRITC

566

150,000

0.04

C5Q

CYAN

Alexa Fluor 546/555
BODIPY TMR
Cy3
DyLight 549
Promo Fluor 546/555
Tetramethylrhodamine (TMR)

550 A

553

Nd YAG

Diode 532 nm green

He-Ne 543 nm green

TRITC

568

150,000

0.04

C5Q

CYAN

Alexa Fluor 546/555
BODIPY TMR
Cy3
DyLight 549
Promo Fluor 546/555
Tetramethylrhodamine (TMR)

555 N (*)

551

Nd YAG

Diode 532 nm green

He-Ne 543 nm green

TRITC

564

77,000

0.70

C5Q

BDP

Alexa Fluor 546/555
BODIPY TMR
Cy3
DyLight 549
Promo Fluor 546/555
Tetramethylrhodamine (TMR)

570 N

573

Krypton 568 nm green

He-Ne 543 nm green

RITC

612

50,000

0.70

C5Q

BDP

Alexa Fluor 568
Cy3.5
Promo Fluor 590
Texas Red

580 N (*)

581

Krypton 568 nm green

He-Ne 543 nm green

RITC

605

75,000

0.70

QSY21

ACEN

Alexa Fluor 568
Cy3.5
Promo Fluor 590
Texas Red

590 A

584

Krypton 568 nm green

He-Ne 543 nm green

RITC

598

125,000

0.14

QSY21

CYAN

Alexa Fluor 568
Cy3.5
Promo Fluor 590
Texas Red

600 N (*)

593

He-Ne 594 nm yellow

Diode 593.5 nm yellow

Texas Red

624

50,000

0.60

QSY21

BDP

Alexa Fluor 594/610
ATTO 594
BODIPY TR
Promo Fluor 60
Texas Red

630 N

629

He-Ne 633 nm red

Diode 635 nm red

 

646

113,000

0.40

QSY21

BDP

Alexa Fluor 633
BODIPY 630/650
Promo Fluor 633

645 C (*)

641

Krypton-Argon 647 nm

Diode 635 nm red

Cy5, APC

663

250,000

0.15÷0.25

QSY21

CYAN

Alexa Fluor 647
Allophycocyanin
BODIPY 650/665
Cy5
DyLight 649
Promo Fluor 647

645 Z (*)

643

Krypton-Argon 647 nm

Diode 635 nm red

Cy5, APC

665

250,000

0.15÷0.25

QSY21

CYAN

Alexa Fluor 647
Allophycocyanin
BODIPY 650/665
Cy5
DyLight 649
Promo Fluor 647

645 A

648

Krypton-Argon 647 nm

Diode 635 nm red

Cy5, APC

667

250,000

0.15÷0.25

QSY21

CYAN

Alexa Fluor 647
Allophycocyanin
BODIPY 650/665
Cy5
DyLight 649
Promo Fluor 647

670 LSS Z (*)

669

He-Ne laser

CyR5.5,
LI-COR 700
channel

778

90,000

0.10

QSY21

 

NIR dye

678 C (*)

679

Krypton 647 nm red

Diode 650 nm red

Diode 670 nm red

CyR5.5,
LI-COR 700
channel

707

240,000

0.10÷0.15

QSY21

CYAN

Alexa Fluor 660/680
Cy5.5
Promo Fluor 680

678 Z (*)

678

Krypton 647 nm red

Diode 650 nm red

Diode 670 nm red

CyR5.5,
LI-COR 700
channel

705

195,000

0.10÷0.15

QSY21

CYAN

Alexa Fluor 660/680
Cy5.5
Promo Fluor 680

678 A

677

Krypton 647 nm red

Diode 650 nm red

Diode 670 nm red

CyR5.5,
LI-COR 700
channel

703

240,000

0.10÷0.15

QSY21

CYAN

Alexa Fluor 660/680
Cy5.5
Promo Fluor 680

690 LSS Z (*)

668

He-Ne laser

CyR5.5,
LI-COR 700
channel

793

90,000

0.10÷0.15

 

 

NIR dye

770 C (*)

678

Xenon-Arc lamp

Dye-pumped

 

790

270,000

0.10

 

CYAN

NIR dye
Promo Fluor 780

770 A

677

Xenon-Arc lamp

Dye-pumped

 

803

270,000

0.10

 

CYAN

NIR dye
Promo Fluor 780

800 C (*)

802

Xenon-Arc lamp

Dye-pumped

NIR,
LI-COR 800
channel

840

250,000

0.10÷0.15

 

CYAN

NIR dye

800 A

802

Xenon-Arc lamp

Dye-pumped

NIR,
LI-COR 800
channel

825

250,000

0.10

 

CYAN

NIR dye

830 C (*)

827

Xenon-Arc lamp
Dye-pumped

 

867

250,000

0.10

 

CYAN

NIR dye

830 A

832

Xenon-Arc lamp

Dye-pumped

 

854

200,000

0.10

 

CYAN

NIR dye

  • COUM: Coumarine
  • BDP: Dipyrromethene borondifluoride
  • ACEN: Acenaphthopyrrole
  • CYAN: Cyanine

NOTE: This is a table of some characteristics of fluorophores useful for Flow Cytometry or Fluorescence Microscopy. Within groups, roughly in order of excitation wavelength (families accepted). Peak excitation and emission wavelengths often vary depending on the environment in which the probe finds itself. Be sure to also look up the excitation and emission spectra for your dye of choice.

 

 

Cyanagen provide the same fluorescent label with different charge values (anionic, cationic, zwitterionic and neutral). It is thus easy to find the most suitable product among the CHROMIS series, starting from the desired absorption/emission peak maximum and subsequently decide both the reactivity and charge properties.

An optimal selection is based on reviewing the spectral properties of each dye, in relation to the instrument to be used. Each instrument has features to consider such as excitation source, optical filters and sensitivity. Therefore, the choice of the right CHROMIS depends on the individual assay as well on the type of instrument.

 

A. Laser excitation source:

Wavelength Color Typical source Suitable CHROMIS dyes
405 nm Blue-Violet Blue-Violet diode laser CHROMIS 425
488 nm Green Argon ion laser CHROMIS 500
514 nm Green Argon ion laser CHROMIS 515
532 nm Green Nd: YAG laser CHROMIS 530
543 nm Green He-Ne laser CHROMIS 550, 555
568 nm Yellow Krypton-ion laser CHROMIS 570, 580
593.5 nm Yellow-Orange Yellow diode laser CHROMIS 600
594 nm Yellow-Orange He-Ne laser CHROMIS 600
633 nm Orange He-Ne laser CHROMIS 630
635 nm Orange Red diode laser CHROMIS 630
647 nm Red Argon-Krypton ion laser CHROMIS 645
650 nm Red Red diode laser CHROMIS 645
670 nm Near IR Red diode laser CHROMIS 678

 

 CHROMIS - Wavelength

Fig 1. Schematic laser sources suitable for CHROMIS fluorescent dyes.

 

B. Continuous white-light excitation source:

Wavelength Color Typical source Suitable CHROMIS dyes
250÷1000 nm UV to IR Xenon-arc lamp All CHROMIS
350÷1000 nm UV to IR Tungsten-halogen lamp All CHROMIS

 

C. Extinction Coefficient (ε): this value is a direct measurement of the dye's ability to absorb light. The ability to absorb light will clearly have an effect on the amount of light it is able to emit.

 

D. Quantum Yield (Φ): the fluorescence quantum yield gives the efficiency of the fluorescence process. It is defined as the ratio of the number of photons emitted to the number of photons absorbed. The maximum fluorescence quantum yield is 1.0 (100%); every photon absorbed results in a photon emitted.

 

E. Charge Value:

Cationic (C): dyes with an intrinsic single positive charge.
Anionic (A): dyes with an intrinsic single negative charge. Highly water soluble and show reduced aggregation allowing easy protein labeling procedures without the need to use organic solvents.
Zwitterionic (Z): dyes with an intrinsic positive and a negative charge.
Neutral (N): dyes with an intrinsic neutral charge.

EXAMPLES OF POST-CONJUGATION APPLICATIONS WITH CHROMIS DYES

GFR measurement (US2015/0328342 A1) GFR measurement (US2015/0328342 A1)
Plasma clearance kinetics in a female Sparague-Dawley rat of ABZWCY-HβCD: 5 mg/100 g body weight by an Electronic Near Infrared Device for Transcutaneous Fluorescence Detection (US 2015/0328342)
Flow Cytometry Flow Cytometry
Expression of human CD4 in lymphocytes from healthy donor in flow cytometry using mouse anti-human CD4 clone SK3 labelled with an OEM Cyanagen dye.
Two dimensional gel electrophoresis Two dimensional gel electrophoresis
Merge of two dimensional electrophoresis analysis between a normal plasma labeled with CHROMIS 645 Z – Iodoacetammide and a plasma from a FSGS (Focal Segmental Glomerulosclerosis) patient labeled with CHROMIS 550 Z – Iodoacetammide. The opposite was done for normal plasma labeled with CHROMIS 550 Z – Iodoacetammide and an FSGS plasma labeled with CHROMIS 645 Z – Iodoacetammide (Proteomics 2009, 9, 460–469).