UE Fluorescence —— Nucleic Acid & Protein Labeling|US Everbright Inc.

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UE Fluorescence —— Nucleic Acid & Protein Labeling

Addtime:2020/5/28 Views:1525 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.

PCR&RT-PCR PCR&RT-PCR

Cell Apoptosis Detection Kit Cell Apoptosis Detection Kit

Cell Viability and Proliferation Assay Kit Cell Viability and Proliferation Assay Kit

TSA immunofluorescence staining TSA immunofluorescence staining

Protein Detection Protein Detection

Molecule Detection Molecule Detection

Cell Staining Reagent Cell Staining Reagent

YF series Fluorescent Dye YF series Fluorescent Dye

Ion and pH Indicator&Enzyme substrate Ion and pH Indicator&Enzyme substrate

Protein and Peptide Labeling Reagent Protein and Peptide Labeling Reagent

Secondary Antibodies Secondary Antibodies

UE Fluorescence —— Nucleic Acid & Protein Labeling

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