Protein Immobilization | Thermo Fisher Scientific

We offer a variety of activated supports and accessories for the immobilization of proteins, antibodies, and other molecules. These resins are available separately or in convenient kits. Choose resins based on NHS-ester, AminoLink™, SulfoLink™, GlycoLink™, CarboxyLink™, and other activation technologies.

Product choice—a variety of coupling chemistries to meet different immobilization strategies
High performance—resins that maximize coupling efficiency and binding capacity
Flexible—available in multiple pack sizes and formats

Choose the right immobilization solution for your experiment


	Pierce NHS-Activated Agarose	AminoLink Coupling Resin	AminoLink Plus Coupling Resin	SulfoLink Coupling Resin	GlycoLink Coupling Resin	CarboxyLink Coupling
Target	-NH2	-NH2	-NH2	-SH	-CHO	-COOH
Support	6% agarose (slurry or dry powder)	4% agarose	4% agarose, highly crosslinked	6% agarose	Ultralink (polyacrylamide/ azlactone copolymer)	4% agarose
Coupling capacity	>25 mg/mL resin	1–20 mg/mL resin	20 mg/mL resin	20 mg/mL resin	1–10 mg/mL resin	5 mg/mL resin
Coupling time	30 min	4 hr	4 hr	2–3.5 hr	30 min	4 hr
Recommended for	Proteins	Proteins, antibodies	Proteins, antibodies	Proteins, peptides, antibodies	Glycoproteins, polyclonal antibodies	Unmodified peptides
Advantages	High coupling efficiency and capacity	High coupling efficiency	High coupling efficiency	Allows for gentle elution	Correctly orients antibodies during coupling	Flexible and gentle coupling conditions
Disadvantages	Narrow pH range (7–9)	May be coupled at antigen-binding site	May be coupled at antigen-binding site	Protein must be reduced first	Not recommended for monoclonal antibodies	Not recommended for antibodies
	26196, 26197 (dry resin), 26200 (slurry)	20381 (10 mL), 20382 (50 mL)	20501 (10 mL), 20505 (50 mL)	20401 (10 mL), 20402 (50 mL), 20404 (250 mL)	53149 (10 mL)	20266 (25 mL)

Thermo Scientific Pierce NHS-Activated Agarose more effectively couples dilute samples than competing products. Rabbit IgG (0.5 mg) was applied to 0.25 mL of activated agarose in PBS at pH 7.2 for 30 minutes. The protein concentration of the flow-through and washes (uncoupled IgG) was determined using the Pierce 660nm Protein Assay. The bound IgG was calculated by subtracting the amount of uncoupled IgG from the amount added. The coupling efficiencies of both dry and slurry forms of Pierce NHS-Activated Agarose were higher than NHS-activated Sepharose™ 4 Fast Flow (GE Healthcare) and Affi-Gel™ 10 Gel (Bio-Rad).

Figure 1. TCEP effectively reduces peptide sulfhydryls (cysteine residues) to maximize covalent immobilization efficiency to SulfoLink Coupling Resin. Peptide A (0.5 mg) and human calcitonin peptide (1 mg) were incubated with (+) or without (-) 25 mM TCEP for 30 minutes and then reacted to 2 mL of SulfoLink Resin for 30 minutes using the default kit procedure. Each peptide contained a C-terminal cysteine, as well as an N-terminal fluorescent tag by which the proportion of non-coupled peptide was determined upon recovery of the reaction solution.

High-yield protein coupling

Immobilization efficiencies for various proteins on 1 mL of AminoLink Plus Resin when immobilized using the pH 7.2 coupling procedure.

Protein	Protein (mg) reacted	Protein (mg) coupled	Coupling efficiency (%)
Protein G	4.6	4.0	83
Mouse IgG	4.7	4.5	96
Rat IgG	4.7	4.4	93
GAM*-IgG	0.9	0.8	84
Human IgG	4.8	4.6	97
Human IgM	0.9	0.8	93

*GAM-IgG is goat anti-mouse IgG, a secondary antibody.

Additional activated supports and accessories are available, as well as convenient kit formats that included disposable columns, buffers, and coupling reagents. Some activated supports are also available in magnetic bead formats.

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For Research Use Only. Not for use in diagnostic procedures.