Sigma-Aldrich Cyanogen bromide-activated-Sepharose™ 4B, lyophilized powder

Cyanogen bromide (CNBr)-activated Sepharose™ 4B is a pre-activated affinity chromatography resin used for covalently coupling ligands containing primary amine groups (such as proteins, peptides, or small molecules) to a Sepharose™ 4B matrix. The matrix is activated with cyanate esters, allowing rapid and efficient coupling under mild conditions, preserving ligand bioactivity.

Supplied as a lyophilized powder stabilized with lactose and dextran, this resin swells when rehydrated to approximately 4–5 mL per gram, making it highly suitable for custom affinity column preparation in research or bioprocess workflows.

Key Features & Benefits

✅ Ready for Ligand Coupling

• Pre-activated with cyanogen bromide for immediate use in coupling primary amine-containing ligands.
• Forms stable isourea or related linkages.

✅ High Binding Capacity with Mild Conditions

• Coupling is performed at neutral to slightly alkaline pH—ideal for preserving protein/ligand activity.
• Typically gives high yields of covalently immobilized ligand.

✅ Versatile and Trusted Affinity Matrix

• Based on Sepharose™ 4B, a well-known, cross-linked agarose bead matrix with excellent flow properties and low non-specific binding.

✅ Flexible Application Scope

• Used in a wide range of applications including:
  • Antibody purification
  • Antigen isolation
  • Protein-protein interaction studies
  • Enzyme immobilization
  • Biomolecule capture assays

✅ Expandable in Scale

• Available in multiple pack sizes (1 g, 5 g, 15 g+) to support both analytical and preparative scale separations.

Product Specifications

Applications

• Affinity Chromatography:
  • Immobilize antibodies, lectins, enzymes, or other ligands for use in specific protein or biomolecule purification.
• Antigen Capture for Immunology:
  • Used in research on xenograft rejection, autoimmune diseases, or vaccine development.
• Protein Purification and Analysis:
  • Purify proteins such as haptoglobin, clusterin, and insecticidal proteins.
• Diagnostics & Biomarker Discovery:
  • Capture target antigens or antibodies from biological fluids like serum or urine.
• Antibody Production and Polyclonal Antibody Purification
  • Frequently used to isolate antibodies from serum or hybridoma supernatants.

Typical Ligand Coupling Protocol (General Guidance)

1. Hydrate 1 g lyophilized resin in 1 mM HCl (4–5 mL expected final volume).
2. Wash thoroughly with coupling buffer (e.g., 0.1 M NaHCO₃, pH 8.3).
3. Add ligand (with free amine groups) and incubate for 2–4 hours at room temperature or overnight at 4°C.
4. Block remaining active groups with 1 M ethanolamine (pH 8.0).
5. Wash with alternating high and low pH buffers to remove unbound material.
6. Store in PBS or Tris buffer with preservative at 2–8°C.

Why Choose Sigma-Aldrich Cyanogen Bromide-Activated Sepharose™ 4B?

✅ Fast, efficient covalent coupling of ligands without complex chemistry

✅ High-capacity, low-background agarose matrix (Sepharose 4B)

✅ Suitable for custom column preparation in immunoprecipitation, pull-down, and purification workflows

✅ Available in flexible pack sizes for R&D or scale-up

✅ Trusted performance from Sigma-Aldrich and Cytiva brands

This product is ideal for researchers working in biochemistry, immunology, proteomics, drug discovery, and molecular biology who require reliable and high-quality chromatography media for custom affinity purification workflows.