Thermo Scientific EZ-Link NHS-PEG Solid-Phase Biotinylation Kit provides a method for amine-targeted antibody biotinylation on-column method for better control of labeling reactions and purification processes.

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

•Suitable for large-scale reactions—1 to 10 mg antibodies
• Quick Labeling and Purification—The entire procedure takes only about an hour
• Easy removal of spent and excess labeling reagents—Only reaction by-products are washed away—no dialysis or Gel filtration
• No dilution effect—Solid phase method allows recovery of initially diluted antibody in smaller volumes after labeling
• Optimized protocol— Antibody-specific protocols ensure appropriate levels of labeling (2 to 5 biotin/antibody molecules), minimizing the possibility of inactivation due to over-labeling
• High Performance Biotin Reagent—NHS -PEG4-Biotin uses highly efficient amine-specific NHS ester chemistry, including hydrophilic polyethylene glycol (PEG) spacers that stabilize long-term solubility of labeled antibodies
• Each kit provides eight bios Biotin labeling experiments—Thermo Scientific No-Weigh packaging ensures the biotin reagent is fully active for eight independent experiments

This innovative antibody labeling system utilizes nickel-chelated agarose to pass through The histidine-rich Fc region temporarily immobilizes the antibody molecule. Once immobilized in place on the resin, the antibody can be biotinylated to the primary amine using a high-quality, solubility-stable NHS-PEG4-biotin reagent. The labeled and purified antibodies are then recovered from the resin using a mild imidazole solution to wash away excess labeling reagents and by-products. No gel filtration or dialysis is required.

EZ-Link Solid Phase Biotinylation Kits use an innovative antibody biotinylation strategy that simplifies many aspects of the process and makes successful biotinylation possible for both beginners and experts. This strategy involves immobilizing the antibody onto a metal-chelated affinity support and performing biotinylation in the solid phase.

Antibodies will readily bind to metal-chelating supports such as nickel or cobalt, primarily through the Fc region of histidine-rich IgG class antibodies. This feature allows for facile derivatization chemistries when the antibody is bound to the support. This strategy facilitates reagent delivery and removal of spent reagents. The resulting modified antibody is then eluted using a buffered imidazole solution to selectively remove the resulting modified antibody from the support.

Solid-phase methods for antibody biotinylation provide better reaction control. For example, reagent and antibody ratios and reaction times can be adjusted to achieve the degree of biotinylation appropriate for the intended application.

This kit contains a water-soluble amino group-specific reagent (NHS-PEG4-biotin) that targets accessible amino (-NH2) groups on the antibody surface. This format can be used at multi-milligram antibody levels in a 1 mL column kit (Product No. 21440) or at low microgram antibody levels suitable for biotinylation using the Micro Spin Column Kit (Product No. 21450).

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