Shop All Antibody Labeling Kits (Conjugation-Based)

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.

DyLight™ 633 Microscale Antibody Labeling Kit (Thermo Scientific™)

The Thermo Scientific DyLight 633 Microscale Antibody Labeling Kit contains an NHS ester-activated derivative of high-performance DyLight 633 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 633 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.

DyLight 633 fluoresces red and has physical properties comparable to other 633 dyes, including Alexa Fluor™ 633. 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 633 Amine-Reactive Dye is available as a stand-alone reagent.

Features of DyLight 633 NHS Ester:

High performance—DyLight 633 shows brighter fluorescence than Alexa Fluor 633
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 633 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 633 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 633 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

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DyLight™ 633 NHS Ester
DyLight™ 633 Antibody Labeling Kit

SAIVI™ Rapid Antibody Labeling Kit, Alexa Fluor™ 750 (Invitrogen™)

The Alexa Fluor® 750 SAIVI™ Antibody Labeling Kit provides a convenient means to label antibodies with an optimal degree of labeling for in vivo imaging applications (DOL; ~2) over a 6-fold antibody concentration range with no adjustments in reaction volume, dye concentration, or antibody concentration necessary. Using this procedure, optimally labeled antibodies are ready for applications that require azide-free reagents, such as live-cell imaging or direct injection into animals.

DyLight™ 650 Microscale Antibody Labeling Kit (Thermo Scientific™)

The Thermo Scientific DyLight 650 Microscale Antibody Labeling Kit is an NHS ester-activated derivative of high-performance DyLight 650 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 650 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.

DyLight 650 provides vibrant far-red fluorescence with comparable or improved performance over other dyes, including Alexa Fluor™ 647 and Cy5™ 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 650 Amine-Reactive Dye is also available as a stand-alone reagent.

Features of DyLight 650 NHS Ester:

High performance—DyLight 650 fluoresces brighter than Alexa Fluor 647 and Cy5
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 650 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 650 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

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DyLight™ 650 NHS Ester
DyLight™ 650 Antibody Labeling Kit

DyLight™ 800 Antibody Labeling Kit (Thermo Scientific™)

Thermo Scientific DyLight 800 Amine-Reactive Dye is an NHS ester-activated derivative of high-performance DyLight 800 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—the amine-reactive DyLight 800 NHS-ester in convenient single-use vials as well as purification resin and spin columns for the preparation of ready-to-use conjugate.

DyLight 800 is a near-IR fluor that is invisible to the naked eye but increases the staining options when using infrared imaging systems. DyLight 800 has spectral properties that are very similar to other near-IR dyes, including Alexa Fluor™ 790 and IRDye™ 800. 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 800 Amine-Reactive Dye is available as a stand-alone reagent or as part of two antibody labeling kit sizes.

Features of DyLight 800 NHS Ester:

High performance— DyLight 800 replaces Alexa Fluor 800 and IRDye 800 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 800 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 800 Amine-Reactive Dye to be provided in multiple formats with flexible protocols to achieve efficient degrees of labeling.

Related Products
DyLight™ 800 NHS Ester
DyLight™ 800 Microscale Antibody Labeling Kit

Alexa Fluor™ 594 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 red fluorescent Alexa Fluor® 594. Comparably small amounts of other proteins (>40 kDa) can also be labeled.

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 Alexa Fluor® 594 Antibody Labeling Kit:
• With an excitation and emission maximum of 590/617 nm, Alexa Fluor® 594 can be efficiently excited using a 594 nm Kr or He-Ne laser line and detected under standard Texas Red® filters
• Labeled proteins typically ready to use typically 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.

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 Zor 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 for use in diagnostic procedures.

SiteClick™ Qdot™ 800 Antibody Labeling Kit (Invitrogen™)

Create a perfectly labeled antibody with the SiteClick™ Qdot® 800 Antibody Labeling Kit. This kit replaces the conventional Qdot® 800 Antibody Conjugation Kit (Q22071MP). 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® 800 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® 800 labels can be used in confocal or traditional fluorescence microscopy.
• In flow cytometry, Qdot® 800 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.

Zenon™ pHrodo™ iFL Green Human IgG Labeling Reagent (Invitrogen™)

Zenon labeling technology provides a fast, versatile, and reliable method for adding a fluorescent label to an antibody. Only a small amount of starting material is needed, and the method is optimized for efficient labeling of antibodies in serum, ascites fluid, or hybridoma suspensions.

Combined with pHrodo iFL dye, this technology provides a rapid, scalable, and easy way to determine how well an antibody internalizes into target cells. pHrodo iFL Green dye dramatically increases its fluorescence as the pH of its surroundings becomes more acidic.

• Labeled antibodies typically ready to use in 10 minutes
• Simple, no purification required
• Fast, accurate internalization results—no need for wash steps or quenchers

Preserve primary antibody function and affinities
Reactive dye labeling of primary antibodies can have unpredictable and undesirable outcomes. One of these is reduced binding affinity due to label addition in the binding pocket. The Zenon antibody labeling approach, which is targeted to the Fc tail, avoids this outcome.

Custom antibody conjugates
If the antibody conjugate you need is not available as one of our stocked products, we will 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.

Pierce™ Conjugate Purification Kit (Thermo Scientific™)

The Thermo Scientific Pierce Conjugate Purification Kit is a gentle affinity chromatography system for purifying horseradish peroxidase (HRP) or alkaline phosphatase (AP) conjugated antibodies from unreacted enzyme after labeling.

Features of the Conjugate Purification Kit:

Specific for IgG—procedure is designed for removal of HRP or AP enzymes from IgG antibodies; not for use with IgM, IgY, Fab fragments or streptavidin conjugates
Complete kit—includes affinity column, activator and elution buffers, desalting column and storage buffers for HRP and AP conjugates
High capacity—the single-use column-kit is designed for purification of 0.5 to 50 mg of antibody conjugate dissolved in Tris-buffered saline
Improves assays—removing unconjugated enzyme from antibody conjugates often reduces background and improves overal performance of immunoassays

This conjugate purification kit is designed to eliminate excess, unconjugated HRP or Alk-Phos enzyme from labeled antibodies (IgG) after performing antibody-enzyme conjugation reactions. The column of metal-chelate agarose resin binds to IgG-enzyme conjugates but allows non-conjugated enzyme (HRP or AP) to to pass through. A gentle, non-denaturing elution buffer recovers the fully active IgG-HRP or IgG-AP conjugate. The included desalting columns exchange the purified conjugate into any buffer of choice, ready for use in various immunodetection methods.

In this kit procedure, the nickel-chelated agarose binds IgG through a histidine-rich cluster on the Fc region at the junctures of the Cγ2 and Cγ3 domains, which is highly conserved across all mammalian IgGs. Purified IgG from sheep, mouse, goat, rat and rabbit will bind to nickel-chelated resin. Avidin or streptavidin enzyme conjugates cannot be purified by this method because they do not bind to the nickel-chelated agarose. Additionally, this kit cannot be used to purify antibodies from serum because other serum proteins will also bind to the nickel column.

Removing non-conjugated enzyme using this kit can improve assay results. Increase in assay sensitivity, shorter washes and incubation times, and a high signal-to-noise ratio can be more easily achieved when free enzyme is absent from the system. Furthermore, the column concentrates samples, which eases assay optimization when using conjugates in dilute solutions.

Alexa Fluor™ 488 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 green fluorescent Alexa Fluor® 488. Comparably small amounts of other proteins (>40 kDa) can also be labeled.

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 Alexa Fluor® 488 Antibody Labeling Kit:
• With an excitation and emission maximum of 494/519 nm, Alexa Fluor® 488 can be efficiently excited using a 488 nm Argon laser line and detected under standard FITC/Cy2® filters
• Labeled proteins typically ready to use typically 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.

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 for use in diagnostic procedures.

SiteClick™ Qdot™ 585 Antibody Labeling Kit (Invitrogen™)

Create a perfectly labeled antibody with the SiteClick™ Qdot® 585 Antibody Labeling Kit. This kit replaces the conventional Qdot® 585 Antibody Conjugation Kit (Q22011MP). 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® 585 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® 585 labels can be used in confocal or traditional fluoresce microscopy.
• In flow cytometry, Qdot® 585 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.

Pacific Orange™ 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 Orange™ has an excitation and emission maximum of 400/551 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.

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

The Thermo Scientific EZ-Link Maleimide-PEG Solid-Phase Biotinylation Kit provides an on-column approach to sulfhydryl-targeted antibody biotinylation that provides greater control over the labeling reaction and clean-up process.

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

Ideal for small-scale reactions—0.1 to 1 mg antibody
Fast labeling and purification—the entire procedure takes only about two hours
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 gentle disulfide reduction and appropriate level of labeling to minimize the possibility of inactivation
High-performance biotin reagent—Malemide-PEG2-Biotin uses efficient, sulfhydryl-specific maleimide 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 gently reduced at the hinge-region to produce free sulfhydryls that are then biotinylated with the Maleimide-PEG2-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 sulfhydryl group-specific reagent (Maleimide-PEG2-Biotin) that targets accessible sulfhydryl (-SH) groups on the surface of the antibody. The format can be applied to low microgram antibody levels for biotinylations using the mini-spin column kit.

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EZ-Link™ Maleimide-PEG Solid-Phase Biotinylation Kit - Prepacked Column

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.

Related Products
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APEX™ Alexa Fluor™ 647 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.