Amine-Reactive Fluorophores, Biotins, Quantum Dots, & Other Labels
Biotin-XX, SE (6-((6-((Biotinoyl)Amino)Hexanoyl)Amino)Hexanoic Acid, Succinimidyl Ester) (Invitrogen™)
The amine-reactive biotin-XX, SE can be used to attach this important hapten to biomolecules of interest for subsequent detection with streptavidin, avidin or NeutrAvidin® biotin-binding protein. This molecule has a 14 atom spacer designated by "XX" to facilitate reactivity with the molecule and eventual accessibility of the biotin to avidin or streptavidin.
DyLight™ 594 NHS Ester (Thermo Scientific™)
Thermo Scientific DyLight 594 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 594 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods.
DyLight 594 provides vibrant red fluorescence with better performance than Alexa Fluor™ 594 and Texas Red™ dye for fluorescent applications. The high water solubility of DyLight Fluors means that a high dye-to-protein ratio can be attained without causing precipitation of the conjugates. DyLight 594 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.
Features of DyLight 594 NHS Ester:
• High performance—DyLight 594 shows brighter fluorescence than Alexa Fluor 594 and Texas Red
• Specific—NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
• Optimized procedure—following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling
Applications:
• Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
• Target protein labeling for in vitro and in vivo fluorescent detection strategies
DyLight 594 Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. Learn more about NHS ester chemistry.
Typical labeling reactions require DyLight 594 Amine-Reactive Dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to specific amounts of the labeling reagent. This feature allows DyLight 594 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.
We also offer Standard and Microscale DyLight 594 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods.The standard size kit contains all necessary components to perform three separate labeling reactions using 1 mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG. Both kit sizes include the Amine-Reactive DyLight 594 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.
Related Products
DyLight™ 594 Antibody Labeling Kit
DyLight™ 594 Microscale Antibody Labeling Kit
DyLight 594 provides vibrant red fluorescence with better performance than Alexa Fluor™ 594 and Texas Red™ dye for fluorescent applications. The high water solubility of DyLight Fluors means that a high dye-to-protein ratio can be attained without causing precipitation of the conjugates. DyLight 594 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.
Features of DyLight 594 NHS Ester:
• High performance—DyLight 594 shows brighter fluorescence than Alexa Fluor 594 and Texas Red
• Specific—NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
• Optimized procedure—following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling
Applications:
• Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
• Target protein labeling for in vitro and in vivo fluorescent detection strategies
DyLight 594 Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. Learn more about NHS ester chemistry.
Typical labeling reactions require DyLight 594 Amine-Reactive Dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to specific amounts of the labeling reagent. This feature allows DyLight 594 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.
We also offer Standard and Microscale DyLight 594 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods.The standard size kit contains all necessary components to perform three separate labeling reactions using 1 mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG. Both kit sizes include the Amine-Reactive DyLight 594 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.
Related Products
DyLight™ 594 Antibody Labeling Kit
DyLight™ 594 Microscale Antibody Labeling Kit
EZ-Link™ NHS-SS-PEG4-Biotin (Thermo Scientific™)
Thermo Scientific EZ-Link NHS-SS-PEG4-Biotin includes disulfide and polyethylene glycol (PEG) groups in its spacer arm to provide biotinylation that is cleavable and which enhances solubility.
Features of EZ-Link NHS-SS-PEG4-Biotin:
• Protein labeling—biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins
• Amine-reactive—reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the amino-termini of polypeptides
• Cleavable—disulfide bond in spacer arm allows the biotin label to be removed using reducing agents such as DTT; only a small sulfhydryl group remains attached to the molecule
• Pegylated—spacer arm contains a hydrophilic, 4-unit, polyethylene glycol (PEG) group
• Enhances solubility—pegylation imparts water solubility to the biotinylated molecule, helping to prevent aggregation of biotinylated antibodies stored in solution
• Long reach—spacer arm (total length added to target) is 37.9 angstroms; this reduces steric hindrance when binding to avidin molecules
NHS-SS-PEG4-Biotin enables simple and efficient biotin labeling of antibodies, proteins and other primary amine-containing macromolecules. The N-hydroxysuccinimide ester (NHS ester) group reacts specifically and efficiently with protein and peptide lysine (K) and N-terminal amino groups at pH 7-9 to form stable amide bonds. The hydrophilic polyethylene glycol (PEG) spacer arm imparts water solubility that is transferred to the biotinylated molecule, thus reducing aggregation of labeled proteins stored in solution. The integral disulfide bond enables the biotin group to be cleaved from the labeled molecule using DTT or other reducing agents, providing for efficient recovery of avidin-bound protein complexes in affinity-based assays.
We manufacture biotin reagents to ensure the highest possible overall product integrity, consistency and performance for the intended research applications.
N-Hydroxysulfosuccinimide (NHS) esters of biotin are the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.
Varieties of biotin NHS-ester reagents differ in length, solubility, cell permeability and cleavability. Non-sulfonated NHS-biotins are cell permeable but must be dissolved in organic solvent such as DMSO or DMF. Sulfo-NHS biotins (and those with pegylated spacers) are directly water soluble but not membrane permeable. Varieties containing disulfide bonds can be cleaved using reducing agents, enabling the biotin group to be disconnected from the labeled protein.
Features of EZ-Link NHS-SS-PEG4-Biotin:
• Protein labeling—biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins
• Amine-reactive—reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the amino-termini of polypeptides
• Cleavable—disulfide bond in spacer arm allows the biotin label to be removed using reducing agents such as DTT; only a small sulfhydryl group remains attached to the molecule
• Pegylated—spacer arm contains a hydrophilic, 4-unit, polyethylene glycol (PEG) group
• Enhances solubility—pegylation imparts water solubility to the biotinylated molecule, helping to prevent aggregation of biotinylated antibodies stored in solution
• Long reach—spacer arm (total length added to target) is 37.9 angstroms; this reduces steric hindrance when binding to avidin molecules
NHS-SS-PEG4-Biotin enables simple and efficient biotin labeling of antibodies, proteins and other primary amine-containing macromolecules. The N-hydroxysuccinimide ester (NHS ester) group reacts specifically and efficiently with protein and peptide lysine (K) and N-terminal amino groups at pH 7-9 to form stable amide bonds. The hydrophilic polyethylene glycol (PEG) spacer arm imparts water solubility that is transferred to the biotinylated molecule, thus reducing aggregation of labeled proteins stored in solution. The integral disulfide bond enables the biotin group to be cleaved from the labeled molecule using DTT or other reducing agents, providing for efficient recovery of avidin-bound protein complexes in affinity-based assays.
We manufacture biotin reagents to ensure the highest possible overall product integrity, consistency and performance for the intended research applications.
N-Hydroxysulfosuccinimide (NHS) esters of biotin are the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.
Varieties of biotin NHS-ester reagents differ in length, solubility, cell permeability and cleavability. Non-sulfonated NHS-biotins are cell permeable but must be dissolved in organic solvent such as DMSO or DMF. Sulfo-NHS biotins (and those with pegylated spacers) are directly water soluble but not membrane permeable. Varieties containing disulfide bonds can be cleaved using reducing agents, enabling the biotin group to be disconnected from the labeled protein.
DyLight™ 425Q NHS Ester (Thermo Scientific™)
Thermo Scientific DyLight 425Q dye is an azo-based dye that can be used to quench matching DyLight and Alexa Fluor dyes. This dye has a λmax of 425 nm (in PBS) and a molar extinction coefficient of 25,000 M-1cm-1. Use this dye with the same laser and filter sets suitable for DyLight 350 and DyLight 405.
General features of DyLight Quencher Specialty Dyes:
• Large selection—the largest family of dyes available for far red-emitting fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
DyLight Quencher Specialty Dyes are a family of non-fluorescent dyes designed for fluorescence quenching in FRET applications. The optimal quencher dye can be selected based upon its characteristic absorption properties that match the emission of a corresponding fluor. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
Criteria to consider when choosing a DyLight Quencher Specialty Dye
• Absorption wavelength—choose the quencher dye with the absorbance wavelength that matches the emission of your fluorophore and the instrument.
• Water solubility—choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight dye selection—the broad selection of red-emitting dyes allows a number of candidate dyes to be tested in an application for optimal performance.
Applications:
• FRET
• Other quenching applications
Related Products
DyLight™ 504Q NHS Ester
DyLight™ 543Q NHS Ester
DyLight™ 641Q NHS Ester
DyLight™ 662Q NHS Ester
DyLight™ 683Q NHS Ester
General features of DyLight Quencher Specialty Dyes:
• Large selection—the largest family of dyes available for far red-emitting fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
DyLight Quencher Specialty Dyes are a family of non-fluorescent dyes designed for fluorescence quenching in FRET applications. The optimal quencher dye can be selected based upon its characteristic absorption properties that match the emission of a corresponding fluor. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
Criteria to consider when choosing a DyLight Quencher Specialty Dye
• Absorption wavelength—choose the quencher dye with the absorbance wavelength that matches the emission of your fluorophore and the instrument.
• Water solubility—choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight dye selection—the broad selection of red-emitting dyes allows a number of candidate dyes to be tested in an application for optimal performance.
Applications:
• FRET
• Other quenching applications
Related Products
DyLight™ 504Q NHS Ester
DyLight™ 543Q NHS Ester
DyLight™ 641Q NHS Ester
DyLight™ 662Q NHS Ester
DyLight™ 683Q NHS Ester
EZ-Link™ NHS-PEG4-Biotin (Thermo Scientific™)
Thermo Scientific EZ-Link NHS-PEG4-Biotin is a pegylated, water-soluble reagent for simple and efficient biotin labeling of antibodies, proteins and other primary amine-containing macromolecules.
Features of EZ-Link NHS-PEG4-Biotin:
• Protein labeling—biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins
• Amine-reactive—reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the amino-termini of polypeptides
• Pegylated – spacer arm contains a hydrophilic, 4-unit, polyethylene glycol (PEG) group
• Enhances solubility – pegylation imparts water solubility to the biotinylated molecule, helping to prevent aggregation of biotinylated antibodies stored in solution
• Irreversible—forms permanent amide bonds; spacer arm cannot be cleaved
• Long reach – spacer arm (total length added to target) is 29 angstroms; this reduces steric hindrance when binding to avidin molecules
NHS-PEG4-Biotin is a long (29.0Å), pegylated, water-soluble, NHS-ester biotinylation reagent to label amines and maximize solubility of antibodies and other proteins. The N-hydroxysuccinimide ester (NHS) group reacts specifically and efficiently with lysine and N-terminal amino groups to form stable amide bonds. The hydrophilic polyethylene glycol (PEG) spacer arm imparts water solubility that is transferred to the biotinylated molecule, thus reducing aggregation of labeled proteins stored in solution. The PEG spacer arm also gives the reagent a long and flexible connection to minimize steric hindrance for binding to avidin molecules.
We manufacture biotin reagents to ensure the highest possible overall product integrity, consistency and performance for the intended research applications.
N-Hydroxysulfosuccinimide (NHS) esters of biotin are the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.
Varieties of biotin NHS-ester reagents differ in length, solubility, cell permeability and cleavability. Non-sulfonated NHS-biotins are cell permeable but must be dissolved in organic solvent such as DMSO or DMF. Sulfo-NHS biotins (and those with pegylated spacers) are directly water soluble but not membrane permeable. Varieties containing disulfide bonds can be cleaved using reducing agents, enabling the biotin group to be disconnected from the labeled protein.
Related Products
EZ-Link™ NHS-PEG4 Biotinylation Kit
EZ-Link™ Micro NHS-PEG4-Biotinylation Kit
Features of EZ-Link NHS-PEG4-Biotin:
• Protein labeling—biotinylate antibodies or other proteins for detection or purification using streptavidin probes or resins
• Amine-reactive—reacts with primary amines (-NH2), such as the side-chain of lysines (K) or the amino-termini of polypeptides
• Pegylated – spacer arm contains a hydrophilic, 4-unit, polyethylene glycol (PEG) group
• Enhances solubility – pegylation imparts water solubility to the biotinylated molecule, helping to prevent aggregation of biotinylated antibodies stored in solution
• Irreversible—forms permanent amide bonds; spacer arm cannot be cleaved
• Long reach – spacer arm (total length added to target) is 29 angstroms; this reduces steric hindrance when binding to avidin molecules
NHS-PEG4-Biotin is a long (29.0Å), pegylated, water-soluble, NHS-ester biotinylation reagent to label amines and maximize solubility of antibodies and other proteins. The N-hydroxysuccinimide ester (NHS) group reacts specifically and efficiently with lysine and N-terminal amino groups to form stable amide bonds. The hydrophilic polyethylene glycol (PEG) spacer arm imparts water solubility that is transferred to the biotinylated molecule, thus reducing aggregation of labeled proteins stored in solution. The PEG spacer arm also gives the reagent a long and flexible connection to minimize steric hindrance for binding to avidin molecules.
We manufacture biotin reagents to ensure the highest possible overall product integrity, consistency and performance for the intended research applications.
N-Hydroxysulfosuccinimide (NHS) esters of biotin are the most popular type of biotinylation reagent. NHS-activated biotins react efficiently with primary amino groups (-NH2) in alkaline buffers to form stable amide bonds. Proteins (e.g., antibodies) typically have several primary amines that are available as targets for labeling, including the side chain of lysine (K) residues and the N-terminus of each polypeptide.
Varieties of biotin NHS-ester reagents differ in length, solubility, cell permeability and cleavability. Non-sulfonated NHS-biotins are cell permeable but must be dissolved in organic solvent such as DMSO or DMF. Sulfo-NHS biotins (and those with pegylated spacers) are directly water soluble but not membrane permeable. Varieties containing disulfide bonds can be cleaved using reducing agents, enabling the biotin group to be disconnected from the labeled protein.
Related Products
EZ-Link™ NHS-PEG4 Biotinylation Kit
EZ-Link™ Micro NHS-PEG4-Biotinylation Kit
DyLight™ 504Q NHS Ester (Thermo Scientific™)
Thermo Scientific DyLight 504Q dye is an azo-based dye that can be used to quench matching DyLight and Alexa Fluor dyes. This dye has a λmax of 504 nm (in PBS) and a molar extinction coefficient of 39,000 M-1cm-1. Use this dye with the same laser and filter sets suitable for DY-415-Co1, DyLight 488 and DEAC.
General features of DyLight Quencher Specialty Dyes:
• Large selection—the largest family of dyes available for far red-emitting fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
DyLight Quencher Specialty Dyes are a family of non-fluorescent dyes designed for fluorescence quenching in FRET applications. The optimal quencher dye can be selected based upon its characteristic absorption properties that match the emission of a corresponding fluor. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
Criteria to consider when choosing a DyLight Quencher Specialty Dye
• Absorption wavelength—choose the quencher dye with the absorbance wavelength that matches the emission of your fluorophore and the instrument.
• Water solubility—choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight dye selection—the broad selection of red-emitting dyes allows a number of candidate dyes to be tested in an application for optimal performance.
Applications:
• FRET
• Other quenching applications
Related Products
DyLight™ 425Q NHS Ester
DyLight™ 543Q NHS Ester
DyLight™ 641Q NHS Ester
DyLight™ 662Q NHS Ester
DyLight™ 683Q NHS Ester
General features of DyLight Quencher Specialty Dyes:
• Large selection—the largest family of dyes available for far red-emitting fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
DyLight Quencher Specialty Dyes are a family of non-fluorescent dyes designed for fluorescence quenching in FRET applications. The optimal quencher dye can be selected based upon its characteristic absorption properties that match the emission of a corresponding fluor. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
Criteria to consider when choosing a DyLight Quencher Specialty Dye
• Absorption wavelength—choose the quencher dye with the absorbance wavelength that matches the emission of your fluorophore and the instrument.
• Water solubility—choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight dye selection—the broad selection of red-emitting dyes allows a number of candidate dyes to be tested in an application for optimal performance.
Applications:
• FRET
• Other quenching applications
Related Products
DyLight™ 425Q NHS Ester
DyLight™ 543Q NHS Ester
DyLight™ 641Q NHS Ester
DyLight™ 662Q NHS Ester
DyLight™ 683Q NHS Ester
6-FAM (6-Carboxyfluorescein), single isomer (Invitrogen™)
The single isomer, 6-FAM, contains a carboxylic acid that can be used to react with primary amines via carbodiimide activation of the carboxylic acid. Fluorescein is the most common fluorescent derivatization reagent for labeling biomolecules. In addition to its relatively high absorptivity, excellent fluorescence quantum yield, and good water solubility, fluorescein has an excitation maximum that closely matches the 488 nm spectral line of the argon-ion laser.
AMCA-X, SE (6-((7-Amino-4-Methylcoumarin-3-Acetyl)amino)Hexanoic Acid, Succinimidyl Ester) (Invitrogen™)
The amine-reactive AMCA-X SE (6-((7-amino-4-methylcoumarin- 3-acetyl)amino)hexanoic acid, succinimidyl ester) and its conjugates yield blue-fluorescence (approximate excitation/emission maxima ~353/442) that can be used as a contrasting color in multicolor applications. Because its fluorescence may not be as bright as that of other dyes or may be obscured by autofluorescence, AMCA is only recommended for use with highly abundant targets. The succinimidyl ester includes and "X" or seven-atom aminohexanoyl spacer between the fluorophore and the reactive group to aid reactivity.
DyLight™ 610-B1 NHS Ester (Thermo Scientific™)
Label peptides, antibodies and other proteins at primary amines with this benzopyrillium-based Thermo Scientific DyLight orange-emitting specialty dye, which is comparable to Alexa Fluor 594, Alexa Fluor 610, Cy3.5 and Texas Red. DyLight 610-B1 dye has excitation and emission peaks at 610 and 632 nm, respectively.
General features of DyLight orange-emitting specialty dyes:
• Large selection—the largest family of dyes available for NIR fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
• Broad spectrum of water solubilities—choose from hydrophilic to hydrophobic dyes to optimize the right dye label for the best performance in a given application
• NIR dyes avoid background interference—DyLight NIR Dyes avoid fluorescence interference or quenching effects from biomolecules present in samples
• Excellent signal penetration through cells and tissues—DyLight NIR Dyes provide the optimal window for excitation and emission for in vivo imaging applications
DyLight NIR Dyes are a family of labeling agents that can be used for bright fluorescence detection in cell-based imaging or in vivo imaging applications. NIR dyes can be selected based upon their characteristic excitation and emission properties or relative hydrophilicity and hydrophobicity attributes. Dyes that contain a greater number of negatively charged sulfonates generally will have greater water solubility than dyes with fewer sulfonates. More hydrophobic dyes often provide better cell penetrating ability in vivo, while more hydrophilic dyes have less nonspecific binding potential. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation. NIR dyes are best for imaging through tissues and away from indigenous fluorescent biomolecule interference or quenching. DyLight Near Infrared Dyes represent the largest selection of fluorescent labels that are commercially available.
Criteria to consider when choosing a DyLight NIR Specialty Dye:
• Excitation and emission wavelengths—choose the best dye to match the excitation and emission capabilities of your instrument
• Water solubility—choose a DyLight NIR Dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight Dye selection—the broad selection of NIR dyes allows a number of candidate dyes to be tested in a given application for optimal performance.
Applications:
• In vivo or ex vivo imaging
• Tumor imaging with labeled peptides
• NIR fluorescence (NIRF) imaging of labeled silica nanoparticles
• NIR in vitro imaging and characterization
• Determination of thermal stability
• Cytotoxicity assays
• Molecular imaging
• UV-VIS-NIR spectroscopy
• Fluorescence correlation spectroscopy
• MRI applications
• DNA sequencing
• Primer labeling for PCR
• 2-D gel electrophoresis
• Flow cytometry/fluorescence-activated cell sorting (FACS)
• Laser scanning confocal microscopy
Related Products
DyLight™ 590-R2 NHS Ester
General features of DyLight orange-emitting specialty dyes:
• Large selection—the largest family of dyes available for NIR fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
• Broad spectrum of water solubilities—choose from hydrophilic to hydrophobic dyes to optimize the right dye label for the best performance in a given application
• NIR dyes avoid background interference—DyLight NIR Dyes avoid fluorescence interference or quenching effects from biomolecules present in samples
• Excellent signal penetration through cells and tissues—DyLight NIR Dyes provide the optimal window for excitation and emission for in vivo imaging applications
DyLight NIR Dyes are a family of labeling agents that can be used for bright fluorescence detection in cell-based imaging or in vivo imaging applications. NIR dyes can be selected based upon their characteristic excitation and emission properties or relative hydrophilicity and hydrophobicity attributes. Dyes that contain a greater number of negatively charged sulfonates generally will have greater water solubility than dyes with fewer sulfonates. More hydrophobic dyes often provide better cell penetrating ability in vivo, while more hydrophilic dyes have less nonspecific binding potential. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation. NIR dyes are best for imaging through tissues and away from indigenous fluorescent biomolecule interference or quenching. DyLight Near Infrared Dyes represent the largest selection of fluorescent labels that are commercially available.
Criteria to consider when choosing a DyLight NIR Specialty Dye:
• Excitation and emission wavelengths—choose the best dye to match the excitation and emission capabilities of your instrument
• Water solubility—choose a DyLight NIR Dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight Dye selection—the broad selection of NIR dyes allows a number of candidate dyes to be tested in a given application for optimal performance.
Applications:
• In vivo or ex vivo imaging
• Tumor imaging with labeled peptides
• NIR fluorescence (NIRF) imaging of labeled silica nanoparticles
• NIR in vitro imaging and characterization
• Determination of thermal stability
• Cytotoxicity assays
• Molecular imaging
• UV-VIS-NIR spectroscopy
• Fluorescence correlation spectroscopy
• MRI applications
• DNA sequencing
• Primer labeling for PCR
• 2-D gel electrophoresis
• Flow cytometry/fluorescence-activated cell sorting (FACS)
• Laser scanning confocal microscopy
Related Products
DyLight™ 590-R2 NHS Ester
Qdot™ 585 ITK™ Amino (PEG) Quantum Dots (Invitrogen™)
Qdot® 585 ITK™ amino (PEG) quantum dots are the ideal starting material for preparing custom conjugates of ultrabright and photostable fluorescently labeled proteins or other biopolymers. These probes are functionalized with amine-derivatized PEG, which prevents non-specific interactions and provides a convenient handle for conjugation. The amino quantum dots react efficiently with isothiocyanates and succinimidyl esters, or with native carboxylic acids using water-soluble carbodiimides such as EDC. Such derivatives may be used for various labeling and tracking applications that require ultrabright and stable fluorescence. Our Qdot® ITK™ amino quantum dots are provided as 8 µM solutions and are available in 8 colors of Qdot® probes.
Important Features of Qdot® ITK™ Amino Quantum Dots:
• Qdot® 585 ITK™ amino quantum dot has emission maxima of ~585 nm
• Extremely photostable and bright fluorescence
• Efficiently excited with single-line excitation sources
• Narrow emission, large stokes shift
• Available in multiple colors
• Ideal for various labeling and tracking applications
Properties of Qdot® Nanocrystals
Qdot® probes are ideal for imaging and labeling applications that require bright fluorescent signals and/or real-time tracking. Unique among fluorescent reagents, all nine available colors of Qdot® probes can be simultaneously excited with a single (UV to blue-green) light source. This property makes these reagents excellent for economical and user-friendly multiplexing applications. Qdot® labels are based on semiconductor nanotechnology and are similar in scale to moderately sized proteins.
About the Innovator’s Tool Kit Qdot® ITK™ Reagents
These Qdot® ITK™ probes are ideal for researchers who wish to prepare specific (non-stocked) conjugates for their applications and need customizable conjugation functionality.
Other Forms of Qdot® Nanocrystals are Available
In addition to the amine-derivatized form, we offer Qdot® ITK™ quantum dots with carboxyl and aliphatic hydrocarbon modifications. We’ve also developed a wide range of Qdot® nanocrystals conjugates and labeling kits. Investigate the properties of Qdot® nanocrystals or read the Molecular Probes® Handbook Section 6.6—Qdot® Nanocrystals to find out more.
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
Important Features of Qdot® ITK™ Amino Quantum Dots:
• Qdot® 585 ITK™ amino quantum dot has emission maxima of ~585 nm
• Extremely photostable and bright fluorescence
• Efficiently excited with single-line excitation sources
• Narrow emission, large stokes shift
• Available in multiple colors
• Ideal for various labeling and tracking applications
Properties of Qdot® Nanocrystals
Qdot® probes are ideal for imaging and labeling applications that require bright fluorescent signals and/or real-time tracking. Unique among fluorescent reagents, all nine available colors of Qdot® probes can be simultaneously excited with a single (UV to blue-green) light source. This property makes these reagents excellent for economical and user-friendly multiplexing applications. Qdot® labels are based on semiconductor nanotechnology and are similar in scale to moderately sized proteins.
About the Innovator’s Tool Kit Qdot® ITK™ Reagents
These Qdot® ITK™ probes are ideal for researchers who wish to prepare specific (non-stocked) conjugates for their applications and need customizable conjugation functionality.
Other Forms of Qdot® Nanocrystals are Available
In addition to the amine-derivatized form, we offer Qdot® ITK™ quantum dots with carboxyl and aliphatic hydrocarbon modifications. We’ve also developed a wide range of Qdot® nanocrystals conjugates and labeling kits. Investigate the properties of Qdot® nanocrystals or read the Molecular Probes® Handbook Section 6.6—Qdot® Nanocrystals to find out more.
For Research Use Only. Not intended for any animal or human therapeutic or diagnostic use.
DyLight™ 488 NHS Ester (Thermo Scientific™)
Thermo Scientific DyLight 488 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 488 for fluorescent labeling of antibodies and other proteins to be used as molecular probes for cellular imaging and other fluorescence detection methods.
DyLight 488 has high fluorescence intensity over a broad pH range (pH 4-9) and is more photostable than Cy2™, Alexa™ Fluor 488, FITC and LI-COR™ dyes in many applications. The high water solubility of DyLight Fluors allows a high dye-to-protein ratio to be achieved without causing precipitation of the conjugates. DyLight 488 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.
Features of the DyLight 488 NHS Ester:
• High performance— DyLight 488 is comparable to Alexa Fluor 488 and brighter than FITC and Cy2
• Specific— NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
• Optimized procedure— following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling
Applications:
• Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
• Target protein labeling for in vitro and in vivo fluorescent detection strategies
DyLight 488 Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. Learn more about NHS ester chemistry.
Typical labeling reactions require the dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to specific amounts of the labeling reagent. This feature allows DyLight 488 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.
We also offer Standard and Microscale DyLight 488 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods. The standard size kit contains all necessary components to perform three separate labeling reactions using 1 mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG. Both kit sizes include the Amine-Reactive DyLight 488 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.
Related Products
DyLight™ 488 Antibody Labeling Kit
DyLight™ 488 Microscale Antibody Labeling Kit
DyLight 488 has high fluorescence intensity over a broad pH range (pH 4-9) and is more photostable than Cy2™, Alexa™ Fluor 488, FITC and LI-COR™ dyes in many applications. The high water solubility of DyLight Fluors allows a high dye-to-protein ratio to be achieved without causing precipitation of the conjugates. DyLight 488 Amine-Reactive Dye is also available as part of two antibody labeling kit sizes.
Features of the DyLight 488 NHS Ester:
• High performance— DyLight 488 is comparable to Alexa Fluor 488 and brighter than FITC and Cy2
• Specific— NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
• Optimized procedure— following the standard protocol results in antibodies with excellent dye:protein ratios and recovery rates for optimum activity and fluorescence labeling
Applications:
• Primary antibody labeling for immunofluorescence microscopy, immunohistochemistry (IHC), Western blotting or ELISA assay
• Target protein labeling for in vitro and in vivo fluorescent detection strategies
DyLight 488 Amine-Reactive Dye is activated with an N-hydroxysuccinimide (NHS) ester moiety to react with exposed N-terminal α-amino groups or the ε-amino groups of lysine residues to form stable amide bonds. Learn more about NHS ester chemistry.
Typical labeling reactions require the dye to first be dissolved in anhydrous dimethyl formamide (DMF) or another suitable organic solvent before adding a specific molar amount of dye to an amine-free buffer containing the protein to be labeled. However, the high solubility of DyLight Fluors permits protein solutions to be added directly to specific amounts of the labeling reagent. This feature allows DyLight 488 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.
We also offer Standard and Microscale DyLight 488 Antibody Labeling Kits for fast and efficient fluorescent labeling of antibodies for use in fluorescence methods. The standard size kit contains all necessary components to perform three separate labeling reactions using 1 mg of IgG or similar quantities of other proteins. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG. Both kit sizes include the Amine-Reactive DyLight 488 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.
Related Products
DyLight™ 488 Antibody Labeling Kit
DyLight™ 488 Microscale Antibody Labeling Kit
DyLight™ 662Q NHS Ester (Thermo Scientific™)
Thermo Scientific DyLight 662Q dye is an benzopyrillium-based dye that can be used to quench matching DyLight and Alexa Fluor dyes. This dye has a λmax of 661 nm (in ethanol) and a molar extinction coefficient of 140,000 M-1cm-1. Use this dye with the same laser and filter sets suitable for DyLight 515-LS.
General features of DyLight Quencher Specialty Dyes:
• Large selection—the largest family of dyes available for far red-emitting fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
DyLight Quencher Specialty Dyes are a family of non-fluorescent dyes designed for fluorescence quenching in FRET applications. The optimal quencher dye can be selected based upon its characteristic absorption properties that match the emission of a corresponding fluor. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
Criteria to consider when choosing a DyLight Quencher Specialty Dye
• Absorption wavelength—choose the quencher dye with the absorbance wavelength that matches the emission of your fluorophore and the instrument.
• Water solubility—choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight dye selection—the broad selection of red-emitting dyes allows a number of candidate dyes to be tested in an application for optimal performance.
Applications:
• FRET
• Other quenching applications
Related Products
DyLight™ 425Q NHS Ester
DyLight™ 641Q NHS Ester
DyLight™ 683Q NHS Ester
DyLight™ 696Q NHS Ester
DyLight™ 766Q NHS Ester
General features of DyLight Quencher Specialty Dyes:
• Large selection—the largest family of dyes available for far red-emitting fluorescence applications
• NHS ester reactive group—allows immediate labeling of antibodies, proteins, peptides and other amine-containing molecules through amide bond formation
DyLight Quencher Specialty Dyes are a family of non-fluorescent dyes designed for fluorescence quenching in FRET applications. The optimal quencher dye can be selected based upon its characteristic absorption properties that match the emission of a corresponding fluor. Each dye contains an amine-reactive NHS ester for simple modification of antibodies, proteins, peptides or other biomolecules through amide bond formation.
Criteria to consider when choosing a DyLight Quencher Specialty Dye
• Absorption wavelength—choose the quencher dye with the absorbance wavelength that matches the emission of your fluorophore and the instrument.
• Water solubility—choose a dye based on its relative hydrophilicity, which directly correlates to the number of negatively-charged sulfonates it has on its core structure. More hydrophilic dyes are best at maintaining water solubility of a labeled antibody and limiting the nonspecific binding of the conjugate. More hydrophobic dyes often are best at penetrating tissues and cell membranes in vivo, meaning that dyes with fewer sulfonates may work best for some applications.
• DyLight dye selection—the broad selection of red-emitting dyes allows a number of candidate dyes to be tested in an application for optimal performance.
Applications:
• FRET
• Other quenching applications
Related Products
DyLight™ 425Q NHS Ester
DyLight™ 641Q NHS Ester
DyLight™ 683Q NHS Ester
DyLight™ 696Q NHS Ester
DyLight™ 766Q NHS Ester
TRITC (5/6-tetramethyl-rhodamine isothiocyanate), mixed isomer (Thermo Scientific™)
Thermo Scientific TRITC is a high-performance derivative of rhodamine dye, activated for easy and reliable labeling of antibodies, proteins and other molecules for use as fluorescent probes.
Features of TRITC:
• Amine-specific labeling—TRITC varieties of rhodamine efficiently label antibodies and other purified proteins at primary amines (lysine side chains)
• Optimized procedure—following the standard protocol results in antibodies with excellent dye:protein ratios for optimum activity and fluorescence
Tetramethylrhodamine isothiocyanate (TRITC) is an amine-reactive derivative of rhodamine dye that has wide-ranging application as antibody and other probe labels for use in fluorescence microscopy, flow cytometry and immunofluorescence-based assays such as western blotting and ELISA.
Applications:
• Label antibodies for use as immunofluorescent probes
• Label oligonucleotides for hybridization probes
• Detect proteins in gels and on western blots
Properties of Rhodamine Dyes:
Thermo Scientific Pierce Rhodamine Dyes are mixtures of isomers with reactive groups attached at the 5- and 6-positions of the bottom ring. The properties of these isomers are indistinguishable in terms of excitation and emission spectra, and for protein applications there is no need to isolate a specific isomer.
TRITC is the base tetramethylrhodamine molecule functionalized with an isothiocyanate reactive group (—N=C=S) at one of two hydrogen atoms on the bottom ring of the structure. This derivative is reactive towards primary amine groups on proteins, peptides and other biomolecules.
Application Data:
Related Products
Pierce™ NHS-Rhodamine Antibody Labeling Kit
NHS-Rhodamine (5/6-carboxy-tetramethyl-rhodamine succinimidyl ester), mixed isomer
Features of TRITC:
• Amine-specific labeling—TRITC varieties of rhodamine efficiently label antibodies and other purified proteins at primary amines (lysine side chains)
• Optimized procedure—following the standard protocol results in antibodies with excellent dye:protein ratios for optimum activity and fluorescence
Tetramethylrhodamine isothiocyanate (TRITC) is an amine-reactive derivative of rhodamine dye that has wide-ranging application as antibody and other probe labels for use in fluorescence microscopy, flow cytometry and immunofluorescence-based assays such as western blotting and ELISA.
Applications:
• Label antibodies for use as immunofluorescent probes
• Label oligonucleotides for hybridization probes
• Detect proteins in gels and on western blots
Properties of Rhodamine Dyes:
Thermo Scientific Pierce Rhodamine Dyes are mixtures of isomers with reactive groups attached at the 5- and 6-positions of the bottom ring. The properties of these isomers are indistinguishable in terms of excitation and emission spectra, and for protein applications there is no need to isolate a specific isomer.
TRITC is the base tetramethylrhodamine molecule functionalized with an isothiocyanate reactive group (—N=C=S) at one of two hydrogen atoms on the bottom ring of the structure. This derivative is reactive towards primary amine groups on proteins, peptides and other biomolecules.
Application Data:
Related Products
Pierce™ NHS-Rhodamine Antibody Labeling Kit
NHS-Rhodamine (5/6-carboxy-tetramethyl-rhodamine succinimidyl ester), mixed isomer
Dapoxyl™ Carboxylic Acid, Succinimidyl Ester (Invitrogen™)
The amine-reactive Dapoxyl® succinimidyl ester can be conjugated to amine-containing biomolecules of interest to create an environmentally sensitive probe. The fluorescence of Dapoxyl® derivatives is very environment sensitive and their fluorescence in water is very low, suggesting that Dapoxyl®conjugates may be useful as probes for conformational changes, denaturation, phosphorylation state and phase changes. Dapoxyl® dye exhibits a huge Stokes shifts (up to ~200 nm) and large extinction coefficient in some solvents.
Oregon Green™ 488 Carboxylic Acid, Succinimidyl Ester, 5-isomer (Invitrogen™)
This single 5-isomer preparation of our proprietary, amine-reactive dye, Oregon Green 488 carboxylic acid, succinimidyl ester and its conjugates have green fluorescence similar to that of fluorescein but are more photostable. Additionally, Oregon Green 488 dye has a lower pKa than fluorescein (pKa = 4.7 versus 6.4 for fluorescein), making this bright dye less pH sensitive in the physiological range.
