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EZ-Link NHS-PEG Solid-Phase Biotinylation Kits


A column-based approach to antibody biotinylation and clean-up.

EZ-Link NHS-PEG Solid Phase Biotinylation Kits

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.

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. Two kits are offered: one for small-scale reactions (0.1 to 1mg antibody) and one for larger scales (1 to 10mg antibody).

Highlights:

  • 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

Product Details:

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 1mL column kit (Part No. 21440) or to low microgram antibody levels for biotinylations using the mini-spin column kit (Part No. 21450).

Protocol scheme for solid-phase biotinylation
Protocol strategy and reaction scheme for biotin labeling with the Thermo Scientific EZ-Link NHS-PEG Solid Phase Biotinylation Kits.

Related Products:

NHS-PEG4-Biotin
Amine-specific Biotinylation Reagents
Maleimide Solid Phase Biotinylation Kits (sulfhydryl-specific)


Ordering Information
 
Product # Description Pkg. Size Instructions MSDS CofA Price
21440 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - 1mL Column
Formulation: Multi-component kit
Sufficient For: 8 labeling reactions, each with 1 to 10mg of antibody

Kit Contents:
Ni-IDA Agarose Column, 1mL, 1 column
PBS pack (makes 500mL), 1 pack
NHS-PEG4-Biotin, 8 x 2mg
Imidazole Stock Solution (4M), 5mL
8-rxn kit Product Instructions for product #21440 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - 1mL Column MSDS for product #21440 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - 1mL Column Certificate of Analysis for product #21440 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - 1mL Column $313.00

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21450 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - Mini-Spin Columns
Formulation: Multi-component kit
Sufficient For: 8 labeling reactions, each with 0.1 to 1mg of antibody

Kit Contents:
Ni-NTA Spin Columns, 0.2mL, 8 columns
PBS pack (makes 500mL), 1 pack
NHS-PEG4-Biotin, 8 x 2mg
Imidazole Stock Solution (4M), 5mL
Collection Tubes (2mL), 30 tubes
8-rxn kit Product Instructions for product #21450 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - Mini-Spin Columns MSDS for product #21450 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - Mini-Spin Columns Certificate of Analysis for product #21450 EZ-Link NHS-PEG Solid-Phase Biotinylation Kit - Mini-Spin Columns $313.00

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Instructions | MSDS | CofA
Product Instructions | MSDS | CofA  

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