Shop All Antibody Labeling Kits (Conjugation-Based)

DyLight™ 755 Microscale Antibody Labeling Kit (Thermo Scientific™)

The Thermo Scientific DyLight 755 Microscale Antibody Labeling Kit contains an NHS ester-activated derivative of high-performance DyLight 755 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection methods. The microscale kit contains all of the necessary components to perform five separate labeling reactions using 100 µg of IgG—the amine-reactive DyLight 755 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.

DyLight 755 is a near-IR fluor that is invisible to the naked eye but increases the staining options when using infrared imaging systems. DyLight 755 has spectral properties that are very similar to other near-IR dyes, including Alexa Fluor™ 750. 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 755 Amine-Reactive Dye is also available as a stand-alone reagent.

Features of DyLight 755 NHS Ester:

High performance—DyLight 755 replaces Alexa Fluor 755 for near-infrared staining
Specific—NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
Convenient kit sizes—standard and microscale sizes are offered to match your experimental needs
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 755 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 755 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

Related Products
DyLight™ 755 NHS Ester
DyLight™ 755 Antibody Labeling Kit

Click-iT™ pHrodo™ iFL Red sDIBO Alkyne for Antibody Labeling (Invitrogen™)

The Click-iT iFL pHrodo Red sDIBO Alkyne for Antibody Labeling is optimized for easy attachment to azido-modified antibodies using copper-free Click chemistry. This sDIBO label can be used with antibodies that have been modified using the SiteClick Antibody Azido Modification Kit or antibodies that have been engineered to contain azido moieties. These sDIBO alkynes are improved versions of our original DIBO cyclooctynes, yielding conjugates that are less "sticky" and give lower signal background in biological samples.

This modular labeling system gives you the option to choose different fluorescent labels for your antibody and attach another molecule via streptavadin or your own molecule via amine-reactive or amine-containing moieties, depending on your assay.

There are multiple Click-iT sDIBO labels to choose from:
Click-iT Alexa Fluor 488 sDIBO Alkyne for Antibody Labeling
Click-iT Alexa Fluor 555 sDIBO Alkyne for Antibody Labeling
Click-iT Alexa Fluor 647 sDIBO Alkyne for Antibody Labeling
Click-iT Biotin sDIBO Alkyne for Antibody Labeling
Click-iT Amine sDIBO Alkyne for Antibody Labeling
Click-iT SDP Ester sDIBO Alkyne for Antibody Labeling

Learn more about SiteClick labeling technology ›

Custom SiteClick Antibody Labeling Service and sDIBO labels
If you have an antibody that is considered "difficult to label" or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor dyes, Qdot fluorophores, R-PE, chelates for PET imaging, and many others.

Pacific Blue™ Antibody Labeling Kit (Invitrogen™)

Molecular Probes™ Antibody Labeling Kits provide a convenient means to label small amounts of antibodies with the violet light-excitable Pacific Orange™ or Pacific Blue™ dyes. This kit is optimized for labeling 100 µg of antibody per reaction. Comparably small amounts of other proteins (>40 kDa) can also be labeled.

View a selection guide for all Antibody Labeling Kits.

View the Fluorophore Selection Guide.

The kit contains everything you need to perform five separate labeling reactions as well as to purify the resulting conjugates. Conjugates are ideal for multiple applications, including flow cytometry, fluorescent microscopy, immunohistochemistry, primary detection, ELISAs, immunocytochemistry, indirect FISH, and more.

Important features of antibody labeling kits:
• Pacific Blue™ has an excitation and emission maximum of 410/455 nm
• Labeled proteins typically ready to use in 90 min (~15 min hands-on time)
• Useful for labeling 100 µg of protein
• Optimized for small-scale labeling of any protein >40 kDa
• Purified using convenient spin filters
• Stabilizing proteins must be removed from the sample before labeling
• Includes detailed instructions for determining degree of labeling (DOL)


Better results and workflows with primary labeled antibodies
A primary antibody directly labeled with a fluorophore often produces lower background fluorescence and less nonspecific binding. Further, multiple primary antibodies of the same isotype or derived from the same species can easily be used in the same experiment if they are directly labeled with compatible fluorophores.

Learn more about protein and antibody labeling
We offer a wide selection of Molecular Probes™ antibody and protein labeling kits to fit your starting material and your experimental setup. See Antibody Labeling from A to Z or use our Labeling Chemistry Selection Tool for other choices. To learn more about our various kits read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes™ Handbook.

We'll make a custom antibody conjugate for you
If you can't find what you're looking for in our stocked list, we'll prepare a custom antibody conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified.

Alexa Fluor™ 790 Antibody Labeling Kit (Invitrogen™)

Molecular Probes® Alexa Fluor® antibody labeling kits provide a convenient means to label small amounts of antibodies with Alexa Fluor® dyes (choice of 10 colors). This kit is optimized for labeling 100 μg of antibody per reaction with infrared fluorescent Alexa Fluor® 790. Small amounts of other proteins (>40 kDa) can also be labeled with this kit.

The Alexa Fluor® 790 Antibody Labeling Kits contains everything you need to perform five separate labeling reactions as well as to purify the resulting conjugates. The conjugates are ideal for multiple applications, including small animal in vivo imaging, flow cytometry, fluorescent microscopy, immunohistochemistry, primary detection, ELISAs, immunocytochemistry, indirect FISH, and more.

With an excitation and emission maximum of 784/814 nm, Alexa Fluor® 790 can be efficiently excited and detected in most standard small animal in vivo imaging and fluorescent western blot imaging systems.

Important Features of Alexa Fluor® 790 Antibody Labeling Kit:

• Labeled proteins typically ready to use in 90 min (~15 min hands-on time)
• Optimized for small-scale labeling of any protein >40 kDa
• Conjugates purified using convenient spin filters
• Stabilizing proteins must be removed from the sample before labeling
• Includes detailed instructions for determining degree of labeling (DOL)

Better Results and Workflows Possible with Labeled Primary Antibodies
A primary antibody directly labeled with a fluorophore often produces lower background fluorescence and less nonspecific binding. Further, multiple primary antibodies of the same isotype or derived from the same species can easily be used in the same experiment if they are directly labeled with compatible fluorophores.

Superior Alexa Fluor® Dyes
Compared to traditional dyes, Alexa Fluor® dyes are brighter, more photostable, and more pH resistant between pH 4 and 10. And generally when using Alexa Fluor® dyes, higher degrees of labeling can be achieved without intramolecular quenching. For details see Alexa Fluor® Dyes Spanning the Visible and Infrared Spectrum—Section 1.3.

Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes® antibody and protein labeling kits to fit your starting material and your experimental setup. See Antibody Labeling from A to Z or use our Labeling Chemistry Selection Tool for other choices. To learn more about our various kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes® Handbook.

We’ll Make a Custom Antibody Conjugate for You
If you can’t find what you’re looking for in our stocked list, we’ll prepare a custom antibody conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified.

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use.

DyLight™ 405 Antibody Labeling Kit (Thermo Scientific™)

The Thermo Scientific DyLight 405 Antibody Labeling Kit contains an NHS ester-activated derivative of high-performance DyLight 405 used to fluorescently label antibodies and other proteins that are then used as molecular probes for cellular imaging and other fluorescence detection 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.

DyLight 405 has a vibrant blue fluorescence intensity that replaces Alexa Fluor™ 405, Cascade™ Blue and AMCA in many 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 405 Amine-Reactive Dye is also available as a stand-alone reagent.

Features of the DyLight 405 Amine-Reactive Dye:

High performance— DyLight 405 shows bright fluorescence and replaces Alexa Fluor 405, Cascade Blue and AMCA
Specific— NHS ester-activated dye labels proteins and other molecules at primary amines (-NH2)
Convenient kit sizes— standard and microscale sizes are offered to match your experimental needs
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 405 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 405 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

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DyLight™ 405 NHS Ester
DyLight™ 405 Microscale Antibody Labeling Kit

EZ-Link™ Plus Activated Peroxidase Kit (Thermo Scientific™)

Thermo Scientific Pierce Plus Activated Peroxidase is an amine-reactive form of horseradish peroxidase (HRP) that provides coupling efficiencies of greater than 95% with antibodies and other proteins. Each kit contains 5 x 1 mg of lyophilized HRP and sufficient Sodium cyanoborohydride solution, quenching buffer, and buffers to conjugate 5 mg of Immunoglobulin (IgG)

Features of the Thermo Scientific Pierce Plus Activated Peroxidase Kit:

Activated HRP – periodate-treated, aldehyde-activated horseradish peroxidase, ready for conjugation to antibodies and other proteins at sites of primary amines (e.g., lysines)
Permanent conjugation – reacts efficiently (95%) with primary amines to form covalent amide bonds upon treatment with sodium cyanoborohydride (included in kit)
High activity HRP – enzyme activity is 120 to 200 units/mg; lyophilized, activated enzyme is stable for at least 12 months at -20°C
Convenient quantities – each 1 mg-quantity of activated enzyme is sufficient for reaction with 1 mg of IgG to produce about 0.5 mL of conjugate
Customizable – vary the molar ratios, reaction buffer and pH, and other parameters to acheive conjugates with different levels of HRP incorporation and activity

Plus Activated Peroxidase is horseradish peroxidase (HRP) enzyme, whose native carbohydrates (sugars) have been gently oxidized with periodate to produce amine-reactive aldehyde groups. These carbonyls of Plus Activated Peroxidase spontaneously and efficiently crosslink with primary amines on antibody or other proteins. The method is more effective than other amine-reactive chemistries, such as glutaraldehyde coupling, which often causes polymerization and a greater degree of conjugate inactivation. The pre-activated enzyme eliminates the difficulties inherent in preparing and validating activated peroxidase from scratch. The kit provides the enzyme, accessory reagents and protocol to easily produce high-performance conjugates for Western blotting, ELISA and other detection techniques.

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EZ-Link™ Plus Activated Peroxidase

APEX™ Alexa Fluor™ 594 Antibody Labeling Kit (Invitrogen™)

The APEX® Antibody Labeling Kits are our best option for covalently attaching a fluorophore to small amounts of IgG antibody (~10–20 μg). It is ideal for the efficient labeling of antibodies in serum, ascites fluid, or hybridoma suspensions. Labeled antibodies are ready for use in imaging or flow cytometry applications in as little as 2.5 hours with very little hands on time.

Important Features of Alexa Fluor® APEX® Antibody Labeling Kits:

• Labeled antibodies typically ready to use in 2.5 hours (~15 minutes hands on time)
• Designed to label 10–20 μg of IgG
• Covalent attachment
• Compatible with contaminating proteins or stabilizers like BSA
• No columns needed; everything you need is supplied for 5 separate labelings
• Choose from Alexa Fluor® 488, 555, 568, 594, and 647 dyes, Oregon Green® 488 dye, Pacific Blue™ dye, and Biotin-XX.


Better Results and Workflows With Primary Labeled Antibodies
A primary antibody directly labeled with a fluorophore often produces lower background fluorescence and less nonspecific binding. Further, multiple primary antibodies of the same isotype or derived from the same species can easily be used in the same experiment if they are directly labeled with compatible fluorophores.

Contaminating Proteins or Protein Stabilizers Are Not a Problem
Many IgG antibodies are often available only in small quantities and packaged with stabilizing proteins, such as BSA, or other contaminants which can interfere with the amine-reactive labeling reagents. The APEX® Antibody Labeling Kits avoids this by utilizing a solid-phase labeling technique that captures the IgG antibody on the resin inside the APEX® antibody labeling tip. Contaminants are simply eluted through the tip, prior to applying the amine-reactive label.

Learn More about Protein and Antibody Labeling
We offer a wide selection of Molecular Probes® antibody and protein labeling kits to fit your starting material and your experimental setup. See Antibody Labeling from A to Z or use our Labeling Chemistry Selection Tool for other choices. To learn more about our various kits read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes® Handbook.

We’ll Make a Custom Antibody Conjugate for You
If you can’t find what you’re looking for in our stocked list, we’ll prepare a custom antibody conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified.

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use.

SiteClick™ Qdot™ 705 Antibody Labeling Kit (Invitrogen™)

Create a perfectly labeled antibody with the SiteClick™ Qdot® 705 Antibody Labeling Kit. This kit replaces the conventional Qdot® 705 Antibody Conjugation Kit (Q22061MP). Unlike the conventional amine-thiol crosslinker method, SiteClick™ labeling specifically attaches the label to the heavy chains of an IgG antibody, ensuring that the antigen-binding domains remain available for binding to your antigen target. This site selectivity is achieved by targeting the carbohydrate domains present on essentially all IgG antibodies regardless of isotype and host species. In addition, no harsh reduction steps are required, and the labeling is consistent and reproducible each time it is performed. Depending upon the label, the resulting SiteClick™ -labeled antibody can be used in flow cytometry, fluorescence imaging, or Western blot detection.

Important Features of the SiteClick™ Qdot® 705 Antibody Labeling Kit:

• Contains everything required to label 100 µg of IgG antibody
• Easy to follow step-by-step protocol
• Highly efficient, site-specific, reproducible labeling chemistry results in high quality antibody conjugate.
• Qdot® 705 labels can be used in confocal or traditional fluorescence microscopy.
• In flow cytometry, Qdot® 705 can be excited by the 488 nm line of the argon-ion laser, or alternatively via excitation at 405 nm. This is true of all Qdot® fluorophores.

Qdot® Fluorophores are Our Brightest Labels
Antibody conjugates made with Qdot® fluorophores produce fluorescence output that surpasses that of traditional organic dyes. Paired with the correct optical filters, Qdot® nanocrystals are as much as 50 times brighter. Read more about Qdot® nanocrystals or review additional product details in Qdot® Nanocrystals—Section 6.6 in the Molecular Probes® Handbook.

Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes® antibody and protein labeling kits (see Antibody Labeling from A to Z). To learn more about our various kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes® Handbook.

Custom SiteClick™ Antibody Labeling Services
If you have an antibody that is considered "difficult to label" or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick™ Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick™ antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor® dyes, Qdot® fluorophores, R-PE, and others.

SiteClick™ Qdot™ 565 Antibody Labeling Kit (Invitrogen™)

Create a perfectly labeled antibody with the SiteClick™ Qdot® 565 Antibody Labeling Kit. This kit replaces the conventional Qdot® 565 Antibody Conjugation Kit (Q22031MP). Unlike the conventional amine-thiol crosslinker method, SiteClick™ labeling specifically attaches the label to the heavy chains of an IgG antibody ensuring that the antigen binding domains remain available for binding to your antigen target. This site selectivity is achieved by targeting the carbohydrate domains present on essentially all IgG antibodies regardless of isotype and host species. In addition, no harsh reduction steps are required, and the labeling is consistent and reproducible each time it is performed. Depending upon the label, the resulting SiteClick™ labeled antibody can be used in flow cytometry, fluorescence imaging, or western blot detection.

Important Features of the SiteClick™ Qdot® 565 Antibody Labeling Kit:

• Contains everything required to label 100 µg of IgG antibody
• Easy to follow step-by-step protocol
• Highly efficient, site-specific, reproducible labeling chemistry results in high quality antibody conjugate.
• Qdot® 565 labels can be used in confocal or traditional fluoresce microscopy.
• In flow cytometry, Qdot® 565 can be excited by the 488 nm line of the argon-ion laser, or alternatively via excitation at 405 nm. This is true of all Qdot® fluorophores.

Qdot® Fluorophores Are Our Brightest Labels
Antibody conjugates made with Qdot® fluorophores produce fluorescence output that surpasses that of traditional organic dyes. Paired with the correct optical filters, Qdot® nanocrystals are as much as 50 times brighter. Read more about Qdot® nanocrystals or review additional product details in Qdot® Nanocrystals—Section 6.6 in the Molecular Probes® Handbook.

Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes® antibody and protein labeling kits (see Antibody Labeling from A to Z). To learn more about our various kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes® Handbook.

Custom SiteClick™ Antibody Labeling Services
If you have an antibody that is considered "difficult to label" or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick™ Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick™ antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor® dyes, Qdot® fluorophores, R-PE, and others.

APEX™ Alexa Fluor™ 555 Antibody Labeling Kit (Invitrogen™)

The APEX® Antibody Labeling Kits are our best option for covalently attaching a fluorophore to small amounts of IgG antibody (~10–20 μg). It is ideal for the efficient labeling of antibodies in serum, ascites fluid, or hybridoma suspensions. Labeled antibodies are ready for use in imaging or flow cytometry applications in as little as 2.5 hours with very little hands on time.

Important Features of Alexa Fluor® APEX® Antibody Labeling Kits:

• Labeled antibodies typically ready to use in 2.5 hours (~15 minutes hands on time)
• Designed to label 10–20 μg of IgG
• Covalent attachment
• Compatible with contaminating proteins or stabilizers like BSA
• No columns needed; everything you need is supplied for 5 separate labelings
• Choose from Alexa Fluor® 488, 555, 568, 594, and 647 dyes, Oregon Green® 488 dye, Pacific Blue™ dye, and Biotin-XX.


Better Results and Workflows With Primary Labeled Antibodies
A primary antibody directly labeled with a fluorophore often produces lower background fluorescence and less nonspecific binding. Further, multiple primary antibodies of the same isotype or derived from the same species can easily be used in the same experiment if they are directly labeled with compatible fluorophores.

Contaminating Proteins or Protein Stabilizers Are Not a Problem
Many IgG antibodies are often available only in small quantities and packaged with stabilizing proteins, such as BSA, or other contaminants which can interfere with the amine-reactive labeling reagents. The APEX® Antibody Labeling Kits avoids this by utilizing a solid-phase labeling technique that captures the IgG antibody on the resin inside the APEX® antibody labeling tip. Contaminants are simply eluted through the tip, prior to applying the amine-reactive label.

Learn More about Protein and Antibody Labeling
We offer a wide selection of Molecular Probes® antibody and protein labeling kits to fit your starting material and your experimental setup. See Antibody Labeling from A to Z or use our Labeling Chemistry Selection Tool for other choices. To learn more about our various kits read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes® Handbook.

We’ll Make a Custom Antibody Conjugate for You
If you can’t find what you’re looking for in our stocked list, we’ll prepare a custom antibody conjugate for you. Our custom conjugation service is efficient and confidential, and we stand by the quality of our work. We are ISO 9001:2000 certified.

For Research Use Only. Not intended for animal or human therapeutic or diagnostic use.

SiteClick™ Qdot™ 525 Antibody Labeling Kit (Invitrogen™)

Create a perfectly labeled antibody with the SiteClick™ Qdot® 525 Antibody Labeling Kit. This kit replaces the conventional Qdot® 525 Antibody Conjugation Kit (Q22041MP). Unlike the conventional amine-thiol crosslinker method, SiteClick™ labeling specifically attaches the label to the heavy chains of an IgG antibody ensuring that the antigen binding domains remain available for binding to your antigen target. This site selectivity is achieved by targeting the carbohydrate domains present on essentially all IgG antibodies regardless of isotype and host species. In addition, no harsh reduction steps are required, and the labeling is consistent and reproducible each time it is performed. Depending upon the label, the resulting SiteClick™ labeled antibody can be used in flow cytometry, fluorescence imaging, or western blot detection.

Important Features of the SiteClick™ Qdot® 525 Antibody Labeling Kit:

• Contains everything required to label 100 µg of IgG antibody
• Easy to follow step-by-step protocol
• Highly efficient, site-specific, reproducible labeling chemistry results in high quality antibody conjugate.
• Qdot® 525 labels can be used in confocal or traditional fluoresce microscopy.
• In flow cytometry, Qdot® 525 can be excited by the 488 nm line of the argon-ion laser, or alternatively via excitation at 405 nm. This is true of all Qdot® fluorophores.

Qdot® Fluorophores Are Our Brightest Labels
Antibody conjugates made with Qdot® fluorophores produce fluorescence output that surpasses that of traditional organic dyes. Paired with the correct optical filters, Qdot® nanocrystals are as much as 50 times brighter. Read more about Qdot® nanocrystals or review additional product details in Qdot® Nanocrystals—Section 6.6 in the Molecular Probes® Handbook.

Learn More About Protein and Antibody Labeling
We offer a wide selection of Molecular Probes® antibody and protein labeling kits (see Antibody Labeling from A to Z). To learn more about our various kits, read Kits for Labeling Proteins and Nucleic Acids—Section 1.2 in the Molecular Probes® Handbook.

Custom SiteClick™ Antibody Labeling Services
If you have an antibody that is considered "difficult to label" or has lost activity after labeling using a conventional method, please contact our custom service representatives to determine whether the SiteClick™ Antibody Labeling Service would be right for your antibody. We offer complete custom SiteClick™ antibody labeling services with the option of multiple detection molecules including biotin, Alexa Fluor® dyes, Qdot® fluorophores, R-PE, and others.

EZ-Link™ NHS-PEG Solid-Phase Biotinylation Kit - Mini-Spin Columns (Thermo Scientific™)

Thermo Scientific EZ-Link NHS-PEG Solid Phase Biotinylation Kits provide an on-column approach to amine-targeted antibody biotinylation that provides greater control over the labeling reaction and clean-up process.

Features of the EZ-Link NHS-PEG Solid-Phase Biotinylation Kit:

Suitable for larger-scale reactions—1 to 10 mg antibody
Fast labeling and purification—the entire procedure takes only about one hour
Easy removal of spent and excess labeling reagent—simply wash away the reaction byproducts—no need for dialysis or gel filtration
No dilution effects—solid-phase method allows initially dilute antibodies to be recovered in a smaller volume after labeling
Optimized protocols—specific protocols for antibody ensure appropriate level of labeling (2 to 5 biotins per antibody molecule), minimizing possibility of inactivation caused by overlabeling
High-performance biotin reagent—NHS-PEG4-Biotin uses efficient, amine-specific NHS-ester chemistry and includes hydrophilic polyethylene glycol (PEG) spacer that stabilizes long-term solubility of labeled antibodies
Eight biotin-labeling experiments per kit—Thermo Scientific No-Weigh Packaging ensures that the biotin reagent is fully active for eight separate experiments

This innovative antibody-labeling system uses nickel-chelated agarose to temporarily immobilize antibody molecules via their histidine-rich Fc regions. Once held in place on the resin, the antibody can be biotinylated at primary amines with the high-quality, solubility-stabilizing NHS-PEG4-Biotin reagent. Excess labeling reagent and byproducts are then washed away before recovering the labeled and purified antibody from the resin using a mild imidazole solution. No gel filtration or dialysis is needed.

EZ-Link Solid-Phase Biotinylation Kits use an innovative strategy for antibody biotinylation that simplifies the process in many respects, making successful biotinylation possible for the novice and expert. The strategy involves immobilizing the antibody to a metal-chelated affinity support and performing the biotinylation in the solid phase.

Antibodies bind readily to metal-chelated supports, such as nickel or cobalt, binding primarily through the histidine-rich Fc region of IgG-class antibodies. This feature allows the derivatization chemistry to be performed easily while the antibody is bound to the support. This strategy facilitates reagent delivery and removal of spent reagent. The resulting modified antibody is then selectively removed from the support by elution with a buffered imidazole solution.

The solid-phase approach to antibody biotinylation provides more control over the reaction. For example: Reagent and antibody ratios and reaction times can be adjusted to achieve the proper degree of biotinylation for the intended application.

The kits contain an aqueous soluble amino group-specific reagent (NHS-PEG4-Biotin) that targets accessible amino (-NH2) groups on the surface of the antibody. The format can be applied at the multi-milligram antibody level in the 1 mL column kit (Part No. 21440) or to low microgram antibody levels for biotinylations using the mini-spin column kit (Part No. 21450).

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EZ-Link™ NHS-PEG Solid-Phase Biotinylation Kit - 1 mL Column