Thermo Scientific™ GlycoLink™ IP Kit

Description

Immobilize polyclonal antibodies or other glycoproteins onto agarose beads for immunoprecipitation or pull-down assays via periodate-oxidized sugars.

The Thermo Scientific GlycoLink IP Kit provides components for effective immunoprecipitation (IP and co-IP) based on small-scale, covalent, affinity-resin immobilization of antibodies and other glycoproteins via oxidized sugar groups.

The GlycoLink Kit uses hydrazide-activated UltraLink Resin to immobilize and perform 25 IP or Co-IP assays with different antibodies or glycoprotein ligands. The method is ideal for polyclonal antibodies, which typically have abundant carbohydrates (glycosylation) on their Fc portions. Monoclonal antibodies that contain adequate carbohydrates are also effective. Antibodies immobilized by this hydrazide-to-carbohydrate method have unobstructed antigen-binding sites and optimal purification capability. The prepared glycoprotein or antibody columns can be regenerated and reused at least five times for immunoprecipitation, co-immunoprecipitation or pull-down affinity-purification without significant loss in binding capacity.

Highlights:

• Efficient immobilization – couple 2 to 10μg of oxidized antibody or other glycoprotein to 20μL of resin in 2 hours or less

• Stable immobilization – resonance structure of the hydrazone bonds are sufficiently stable so the antibody remains coupled during elution

• Preserves binding function – immobilizes IgG via the Fc region, thereby keeping both antigen binding sites available for capturing target

• Versatile applications – immobilize any molecule that contains oxidizable sugars, including glycoproteins for use in purifying protein interaction binding partners

Includes:

Kit contains resin, buffers and reagents for immobilization; desalting columns; IP lysis/wash and elution buffers for immunoprecipitation; microcentrifuge spin columns and collection tubes; and loading buffer for SDS-PAGE

Recommended for:

Immunoprecipitation followed by electrophoresis and excision of the protein for mass spectrometry or sequencing; Immunoprecipitation and SDS-PAGE analysis of a protein with a molecular weight similar to the antibody's heavy or light chain; Identification of binding partners of immobilized glycoprotein

Co-immunoprecipitation for determining protein-protein interactions

Specifications

• Capacity (Metric): 2 to 10 μg Oxidized Antibody or Glycoprotein Coupling to 20 μL Resin
• Content And Storage: Sufficient For: 25 IP/Co-IP reactions, each using 20 μL antibody-coupled resin
  • UltraLink Hydrazide Resin, 0.5 mL
  • GlycoLink Coupling Buffer, 50 mL
  • Aniline, 0.2 mL
  • Sodium meta-periodate, 0.5 g
  • Wash Buffer, 60 mL
  • Zeba Spin Desalting Columns,0.5 mL, 25 columns
  • IP Lysis/Wash Buffer, 50 mL
  • PBS Pack (makes 500 mL), 1 pack
  • Elution Buffer, pH 2.8, 50 mL
  • Lane Marker Sample Buffer, Non-reducing, (5X), 5 mL
  • Pierce Spin Columns-Screw Caps, 25 columns
  • Collection Tubes, 2 mL, 100 tubes
  
  Store at 4°C.
• Assay: Affinity Purification
• For Use With (Equipment): Microcentrifuge
• Product Line: GlycoLink™
• Quantity: 25 reactions kit
• For Use With (Application): Immunoprecipitation (IP)
• Target: Not Target-Specific
• Type: Immunoprecipitation/Co-Immunoprecipitation Kit
• Sufficient For: 25 Reactions
• Format: Kit

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