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UE Fluorescence —— Nucleic Acid & Protein Labeling

Addtime:2020/5/28 Views:1657 Sources:US Everbright Inc.

US Everbright® Inc. (UE) provides multiple fluorescent synthetic dyes: YF series (350-790 nm), Cy series and probe-synthetic dyes, etc.


1. YF series - Nucleic Acid & Protein Labeling

(1)  Origin of YF dyes



YF is a series of highly water-soluble fluorescent dyes. As a next generation of protein- and nucleic-acid-labeling dyes, YF is equivalent to the products like Alexa fluor, dylight, Atto and iFluor. YF series have incomparable brightness and optical stability, excellent water solubility, and more than 20 kinds of products.

Figure 1 Spectral characteristics and fluorescence distribution of YF dyes


(2) Features of YF dyes

Excellent light stability, difficult to bleach, suitable for microscopy

Great water solubility, and insensitive to pH

▪ High fluorescence quantum yield,high fluorescence brightness (≥4), high SNR, specific for the detection of expressed proteins

High specificity,and offering different proteins a variety of dyes with different wavelengths to improve their positive expression.


(3) Applications of YF dyes

①Labeling of proteins, nucleic acids and other molecules 

Form

Application

Carboxylic acid/Free acid

Inactive form

Amine-modified imine succinate (SE, water soluble/liposoluble)

Label free amino groups of target molecules, such as lysine residues

Mercaptan-modified Maleimide (MTS)

Label sulfhydryl groups of target molecules, such as cysteine residues. MTS can replace SE to modify dyes, in the case that SE interferes with the biological activity of the protein,

Aminooxy

No reductant is needed to react with aldehyde or ketone group to generate a stable connection

Hydrazide

Label aldehyde or ketone groups of target molecules. Hydrazides of YF dyes can also be used as tracers to study the morphology and gap junction of neurons.

Amine

React with carboxylic groups activated by biomolecules。


②YF Biomarkers

YF Biomarkers

Application

Relevant Products

Annexin V

Cell apoptosis (phosphatidylserine)

Annexin V Kit

α-Bungarotoxin

Probe of acetylcholine receptor

——

Bovine serum albumin

Tracer of blood flow in vivo

——

Concanavalin A

Carbohydrate probe (agglutinins)

——

dUTP

DNA probe,TUNEL assay

dUTP labeling, TUNEL Kit

Azide and Picolyl azide

Cell-Proliferation Assay of EDU

EDU Kit

Phalloidin

F-actin probe

Phalloidin Kit

Strepatavidin

Biotin detection

——

Wheat germ agglutinin

Carbohydrate probe (agglutinins), Gram staining

——

DBCO/BCN, Alkyne

Reaction with azide

——

Tyramide (Tyramine signal amplification, TSA)

High density situ labeling of target proteins or nucleic acids

with horseradish peroxidase (HRP), based on enzyme detection method.

HRP chromogenic Kit


(4) Profile of YF dyes

YF dyes

λEX (nm)

λEM (nm)

Excitation wavelength

Substitutes

Features and applications

YF350

347

448

UV

AF350, AMCA, DyLight 350

• Brighter blue fluorescence

• Good water solubility, insensitive to pH

YF405S

404

431

405 nm

AF405, Cascade Blue, DyLight 405

• Flexible for most instruments

YF405M

408

452

405 nm

BD Horizon V 405,eFluor  450,Pacific Blue

• Stable fluorescence

• Specific for super-resolution fluorescence imaging of SIM**

YF488A

490

515

488 nm

ATTO 488, AF488, Cy2, DyLight 488, FAM, FITC, Fluorescein

• Less non-specific binding and less fluorescence overflow than Alexa Fluor 488

• Stable fluorescence

• Specific for super-resolution fluorescence imaging of TIRF**

YF543

541

560

532, 543, 546 nm

AF546, TAMRA

• Brighter than Alexa Fluor 546

YF555

555

565

532, 543, 546, 555, 568 nm

AF555, ATTO550, Cy3, DyLight 549, TRITC

• Brighter than Cy3

• Specific for super-resolution fluorescence imaging of STORM**

YF568

562

583

532, 543, 546, 555, 568 nm

AF568, ATTO565, Rhodamine Red, R-PE

• Brighter than Alexa Fluor 568

• Applicable for fluorescence imaging of TIRF and multicolor STORM**

YF594

593

614

532, 543, 546, 555, 568 nm

AF594, ATTO594, DyLight 594, Texas Red,  Cy3.5

• Generate the brightest conjugates among dyes with similar spectra

• Super stable fluorescence

YF620R

617

639

633, 635 nm

LightCycler Red 640

• With unique fluorescence spectrum and brighter fluorescence

YF633

630

650

633, 635 nm

AF633, AF647, Cy5, DyLight 633

• Generate the brightest antibody-conjugates among dyes with similar spectra

• More stable than Alexa Fluor 647

• Specific for super-resolution fluorescence imaging of TIRF, FIONA and gSHRImP**

YF640R

642

662

633, 635, 640 nm

AF647, ATTO 647N, Cy5, DyLight 649

• More stable than Cy5

• Generate brighter protein conjugates

• Applicable for super-resolution fluorescence imaging of TIRF and FLIMP

YF647

650

665

633, 635, 640 nm

AF647, ATTO 647N, Cy5, DyLight 649

• Brighter than Cy5

• Applicable for super-resolution fluorescence imaging of STORM**

YF660C

667

685

633, 635, 640 nm

AF660

• Brighter than Alexa Fluor 660

• Applicable for super-resolution fluorescence imaging of STORM**

YF660R

663

682

633, 635, 640 nm

AF660, APC, Cy5.5

• Brighter than Alexa Fluor 660

• Super stable fluorescence

YF680

681

698

680, 685 nm

AF680, Cy5.5, DyLight680, IRDye 680LT

• Brighter than fluorescent dyes with similar spectrums

• Applicable for LI-COR® Odyssey® System

YF680R

680

701

680, 685nm

AF680, Cy5.5, DyLight680, IRDye 680LT

• Brighter and more stable fluorescence

• Applicable label nucleic acids and other small molecules

• Applicable for LI-COR® Odyssey® System

• Applicable for STED and single molecule spectroscopy**

YF750

755

777

680, 685nm

AF750, Cy7, DyLight750, APC-AF 750, IRDye 750

• Brighter and more stable fluorescence

• Good water-solubility

• Applicable for super-resolution fluorescence imaging of STORM**

YF770

770

797

785nm

DyLight 800, IRDye 800CW

• Brighter and more stable fluorescence

• Applicable for LI-COR® Odyssey® System

YF790

784

806

785nm

AF790

• Brighter and more stable fluorescence

• Good water solubility

UE also provides LF series labeling antibodies and nucleic acids, such as LF650.


2. Cy dyes

Cyanine Dyes (Cy Dyes) are a class of fluorescent dye used for labeling amine groups on antibodies, peptides or oligonucleotides, and are suitable for use in a range of applications, including microscopy, flow cytometry and fluorescence resonance energy transfer (FRET).

(1) Applications of Cy dyes

•Labeling of soluble proteins (resisting to organic co-solvent)

•Antibody labeling (dissolved in 5-10% DMSO / DMF)

•DNA and oligo labeling

•polypeptide labeling

•labeling of other molecules

(2) Categories of Cy dyes

UE provides various Cy dyes, like water-soluble/water-insoluble and methyl (M)-/ ethyl (E)-modified forms.

Water-soluble ethyl-modified(E)series is compared to Quasar series,water-insoluble methyl-modified(M)series is compared to Lumiprobe series

(3) Profile of Cy series


Cy3

Cy3.5

Cy5

Cy5.5

Cy7

Cy7.5

Absorbance max

550 nm

581nm

649 nm

675 nm

747 nm

788 nm 

Extinction max

150 000 M-1cm-1

150 000 M-1cm-1

250 000 M-1cm-1

250 000 M-1cm-1

200 000 M-1cm-1

 223000 M-1cm-1

Emission max

570 nm

596 nm

670 nm

694 nm

776 nm

 808 nm


3. Common probe-synthetic dyes

UE provides plenty of common probe-synthetic dyes, such as Rox, Fam, Tamra, FITC and Quasar. 


    

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