CNBr-activated Agrose - BioReagent,50% v/v; 90μm

Cat. No.: C1523223

GRADE & PURITY BioReagent, 50% v/v; 90μm

Storage

Store at 2-8°C,Do not freeze

Shipped In

Wet ice

Application

Protein purification

Übersicht

CNBr-activated Agrose is a pre-activated agarose medium. The matrix is 4% highly cross-linked agarose with good physical and chemical stability. This medium is commonly used for coupling antibodies or high-molecular-weight proteins containing primary amino groups. The coupling reaction is spontaneous and rapid, and the coupled medium exhibits stable affinity properties. It is mainly used for the conjugation and immobilization of proteins, peptides and nucleic acids.

Aladdin CNBr-activated Agrose is stored in acetone at a 1:1 volume ratio of gel to protective solution. The product specification refers to the volume of the actual gel.

Parameter	Specification
Matrix	4% Agarose
Activated group	Cyanate ester
Average particle size	90 μm (45–165 μm)
Coupling conditions	pH 8–10, room temperature (20–25 °C) for 2–4 h or 4 °C overnight
Coupling capacity	≥15 mg BSA/mL wet gel
Recommended flow rate	<75 cm/h
Working pH range	3–11
CIP pH range	2–11
Chemical stability	Stable in common aqueous buffers; compatible with denaturants such as 8 M urea or 6 M guanidine hydrochloride
Storage conditions	100% acetone; 2–8 °C
Shelf life	2 years

Instructions for Use

To ensure full activity, this product is stored in 100% acetone at 2–8 °C for long-term storage. Short-term transportation and use are allowed at room temperature.

1.Solution Preparation

Buffer	Volume
Solution A: 1 mM HCl, 0.5 M NaCl (pre-chilled, pH 2–3)	≥20× medium volume
Solution B: 0.2 M NaHCO<sub>3</sub>, 0.5 M NaCl, pH 8.3	≥10× medium volume
Solution C: 0.1 M Tris, pH 8.0	≥5× medium volume
Solution D: 0.05 M Tris, 0.5 M NaCl, pH 8–9	≥15× medium volume
Solution E: 0.1 M acetate buffer, 0.5 M NaCl, pH 3–4	≥15× medium volume
Solution F: 0.02 M PB, 0.15 M NaCl, pH 7.0–7.6	≥10× medium volume

2.Ligand Pretreatment

Typically, 1 mL medium can couple approximately 5–10 mg protein ligand.

Powdered protein ligand: dissolve in Solution B.

Liquid protein ligand: exchange buffer to Solution B before use.

3.Medium Pretreatment

Place an appropriate amount of affinity chromatography medium (stored in acetone) in a glass sintered funnel. Wash with 15× gel volume of pre-chilled Solution A.

4.Ligand Coupling

After washing, transfer the medium into the ligand solution at a 1:1 volume ratio and mix well.ncubate on a shaker at room temperature for 2–4 h, or at 4 °C overnight.

5.Washing

After reaction, wash with 5× medium volume of Solution B to remove uncoupled protein ligand.

6.Blocking

Wash the medium with 3× medium volume of Solution C, then block remaining activated sites with Solution C at a 1:1 volume ratio.Incubate on a shaker at room temperature for 2–4 h.

7.Alternate Washing

After blocking, wash alternately with Solution D and Solution E for 3–6 cycles, using 3× medium volume per wash.

8.Storage

Finally, wash with 10× medium volume of Solution F (other binding buffers may be used as needed).Ligand coupling is complete and the affinity medium is ready for use.For long-term storage, store the medium in 20% ethanol at 2–8 °C.

Factors Affecting Coupling

1.Coupling BufferBicarbonate or borate buffers are commonly used. Do not use Tris or other amino-containing buffers as coupling buffers.To maintain high coupling efficiency, pH is usually 8–10.To reduce protein–protein adsorption, add an appropriate salt concentration (e.g.0.5 M NaCl).

2.Temperature and Reaction TimeCoupling is completed within 2–3 h at room temperature.For low-temperature coupling (e.g.4 °C), incubate overnight (10–16 h).

3.Ligand ConcentrationRecommended loading: 5–10 mg protein ligand per mL medium.Excess ligand may cause high ligand density, leading to steric hindrance, reduced binding capacity, difficult elution, and increased non-specific adsorption.

Note: Low pH reduces coupling activity. However, to minimize steric hindrance or preserve ligand bioactivity, coupling may be performed at lower pH (e.g.pH 6).

Regeneration

A typical regeneration procedure:Wash alternately with high-pH buffer (e.g.0.1 M Tris, 0.5 M NaCl, pH 8.5) and low-pH buffer (e.g. 0.1 M NaAc, 0.5 M NaCl, pH 4.5) for 3–5 cycles, using 2–3× medium volume per wash.Finally equilibrate with binding buffer.

Note: Regeneration varies by ligand; this method is for reference only.

Cleaning

The medium requires cleaning after 3–5 cycles to maintain performance.

Recommended cleaning agents for different contaminants:

Microorganisms: NaCl, filter-sterilized buffers, non-ionic detergents.

Tightly bound proteins: pH 8.5 buffer with non-ionic detergent or 20%–70% ethanol.

Storage Conditions

2–8 °C, 100% acetone

Specifications

Spezifikationen & Reinheit

BioReagent,50% v/v; 90μm

Stabilität und Lagerung

Store at 2-8℃. long term (24months).

Storage

Store at 2-8°C,Do not freeze

Verschickt in

Wet ice

Dieses Produkt erfordert Kühlkettenversand. Grundversand und andere Economy-Optionen sind nicht verfügbar.

Note

BioReagent

Product Line

Biologische Produkte

Application Protocols

General workflow protocols for using CNBr-activated agarose (literature-based; adjust to your ligand and scale):

A. Ligand Coupling and Blocking

1. Wash 1 mL settled resin with 5 mL cold 1 mM HCl (or DI water) in a fritted column; drain completely.
2. Equilibrate with 5 mL coupling buffer (0.1 M NaHCO3, 0.5 M NaCl, pH ~8.3).
3. Apply ligand (e.g., 2–10 mg in 1–5 mL coupling buffer) and incubate 2 h at RT or overnight at 4 °C with gentle mixing. Collect flow-through to estimate coupling efficiency (A280 or relevant assay).
4. Wash with 5–10 CV coupling buffer.
5. Block with 1 M ethanolamine, pH ~8.0 for 1–2 h at RT; then wash alternately with low-pH acetate buffer (0.1 M, 0.5 M NaCl, pH ~4) and high-pH Tris buffer (0.1 M, 0.5 M NaCl, pH ~8–9), 3 cycles each.

B. Column Packing and Equilibration

1. Slurry-pack into an appropriate column using equilibration buffer suited to the immobilized ligand (e.g., PBS, pH 7.4). Avoid high pressures; monitor bed height and clarity.
2. Equilibrate with 5–10 CV of running buffer.

C. Affinity Capture (example for protein targets)

1. Load clarified sample at 0.5–1 CV/min (gravity or low-pressure). Collect flow-through for mass balance.
2. Wash with 10 CV running buffer until baseline UV.
3. Elute with ligand-appropriate conditions (e.g., low pH glycine pH 2.5–3.0, high salt, or competitive ligand). Immediately neutralize low-pH fractions if needed for protein stability.

D. Regeneration and Storage

1. Wash with 5–10 CV running buffer; apply brief low/high pH wash if fouled.
2. Store at 2–8 °C in a suitable preservative-containing buffer (if compatible). For this item, specific preservative content is not specified; refer to CoA/Spec Sheet.

Biological Roles

This product is a research chromatographic matrix rather than a biological macromolecule with intrinsic cellular function. The following notes provide context about the base material (agarose) and its compatibility in biochemical workflows (general/literature):

• Agarose origin and nature: Agarose is a neutral polysaccharide derived from red seaweed (Rhodophyta). It forms thermo-reversible gels through hydrogen bonding and helix formation. As a chromatographic matrix, it is typically crosslinked for mechanical stability.
• Biochemical compatibility: The matrix is largely inert, nonionic, and exhibits low nonspecific protein binding compared to many synthetic polymers, making it suitable for protein immobilization and affinity capture.
• Reactivity: CNBr activation appends reactive groups on the agarose surface; these groups are designed to react with ligand amines, not to elicit biological responses. After coupling and blocking, the surface becomes largely unreactive aside from the immobilized ligand’s function.
• Enzyme immobilization: Agarose’s hydrophilicity can preserve native-like hydration shells around immobilized enzymes, often supporting retained activity and allowing repeated use under gentle conditions (outcome is enzyme- and protocol-dependent).
• Diffusion and mass transport: The porous structure supports diffusion of proteins up to hundreds of kDa depending on pore size (resin-specific; not specified for this item). For very large complexes, lower flow rates improve binding by reducing mass-transfer limitations.

No clinical or in vivo biological roles are claimed or implied. This product is for research use only (from Product Data).

Buffer Applications

While this product is not itself a buffer, successful use depends on appropriate buffer systems for coupling, washing, and elution (general/literature guidance):

• Coupling buffer (amine-targeted):
  • Typical recipe (literature): 0.1 M NaHCO3, 0.5 M NaCl, pH ~8.3. The salt reduces nonspecific ionic adsorption; bicarbonate provides mildly basic pH to favor deprotonation of primary amines.
  • Avoid amine-containing buffers (Tris, glycine, ammonium) and additives with primary amines during coupling.
• Ligand preparation: Exchange the ligand into coupling buffer (e.g., by desalting column or dialysis). Maintain ligand concentration sufficient for near-saturation of reactive groups while avoiding precipitation.
• Blocking solution: 1 M ethanolamine, pH ~8.0 (literature) to quench unreacted active groups after coupling.
• Wash regimen (desorption of noncovalent binders): Alternate between low pH (e.g., 0.1 M acetate, 0.5 M NaCl, pH ~4) and high pH (e.g., 0.1 M Tris-HCl, 0.5 M NaCl, pH ~8–9) several cycles to remove adsorbed ligand and impurities (avoid amine buffers before blocking).
• Storage buffer (post-coupling): Commonly PBS or appropriate neutral buffer containing a microbial preservative such as 20% ethanol or 0.02% sodium azide (literature). For this item, any specific preservative content is not specified; confirm compatibility with your application and safety policies.
• Operational buffers for affinity use: Chosen based on the immobilized ligand-target pair (ionic strength, pH, cofactors). For antibody affinity, isotonic buffers with gentle detergents can reduce nonspecific binding.

All numeric specifications (capacities, exact tolerances) are not specified for this item; consult CoA/Spec Sheet.

Green Alternatives

Perspective on greener choices for affinity matrix activation (general/literature):

• Environmental/safety context:

  • CNBr activation uses and historically required handling of toxic cyanogen bromide. Purchasing pre-activated agarose (as in this product) mitigates direct handling but the activation chemistry still bears inherent hazards upstream.

• Potential alternatives and trade-offs:

  • NHS-activated agarose:
    • Pros: Highly efficient amine coupling at near-neutral pH, well-defined succinimidyl ester chemistry; often higher reproducibility.
    • Cons: NHS esters are moisture-sensitive with finite shelf life; can be costlier.
  • Epoxy-activated agarose:
    • Pros: No cyanide-related chemistry; can couple to a variety of nucleophiles (amines, thiols, hydroxyls) under alkaline conditions.
    • Cons: Requires longer coupling times and higher pH/temperature, potentially harsher on sensitive ligands.
  • Carboxylated agarose with EDC/NHS in situ:
    • Pros: Avoids CNBr; zero-length coupling to amines.
    • Cons: Requires handling water-soluble carbodiimides; more steps and byproduct management.
  • Thiol-directed chemistries (e.g., maleimide- or vinylsulfone-activated agarose):
    • Pros: Site-selective coupling to cysteines; can preserve ligand activity.
    • Cons: Requires engineered thiols; maleimides can hydrolyze and undergo exchange.

• Operational green tips (applicable regardless of chemistry):

  • Work at ambient temperature when possible; minimize energy use by avoiding prolonged refrigerated incubations if ligand stability allows.
  • Use buffered saline systems and minimize organic solvents for cleaning; validate lower-concentration CIP agents.
  • Regenerate and reuse immobilized columns to reduce waste; document resin lifetime.

No item-specific environmental certifications or green metrics are provided; refer to CoA/Spec Sheet.

Pharmaceutical Uses

This product is labeled For research use only and is not intended for clinical manufacturing or diagnostic use. The following remarks are for general context (no claims):

• Role in development: CNBr-activated agarose is commonly used in R&D to prototype custom affinity resins (e.g., enzyme immobilization, antibody capture) that, if successful, may later be translated to qualified/GMP-grade materials for bioprocessing.
• Excipient status: Agarose matrices are not pharmaceutical excipients per se when used as solid-phase media; pharmacopeial monographs typically address agar/agarose as analytical reagents or for electrophoresis. No pharmacopeial status is claimed for this item.
• Process relevance: Affinity resins prepared on CNBr-activated agarose can support downstream purification method development, host-cell protein depletion studies, or analytical enrichment prior to LC–MS.
• Scalability considerations (general): For potential scale-up, one would require defined bead size, porosity, pressure-flow characteristics, ligand coupling capacity, and validated clean-in-place/sanitization procedures—parameters not specified for this item and typically documented for process-grade media.
• Regulatory note: Any use in cGMP environments requires appropriate supplier qualification, change control, extractables/leachables data, and lot-specific CoA. This research-grade product is not represented as GMP-compliant.

Item-specific claims such as endotoxin limits, bioburden controls, or viral safety are not provided; refer to CoA/Spec Sheet or inquire for process-grade alternatives if needed.

Physical Properties

Item-specific physical data:

• Appearance: Not specified for this item; refer to CoA/Spec Sheet. (Typically supplied as a hydrated bead slurry, general note only.)
• Density, particle size, porosity, water content: Not specified for this item; refer to CoA/Spec Sheet.
• pH of storage solution/preservatives: Not specified for this item; refer to CoA/Spec Sheet.

Polymer/resin general properties (literature/general):

• Physical state: Hydrated, porous gel beads (insoluble in water and common organic solvents; swells in aqueous media).
• Solubility: Insoluble; beads are suspended in aqueous buffers. Compatible with most aqueous salt solutions and many denaturing agents (e.g., urea, guanidinium) within typical concentration limits for agarose resins.
• Stability of activation: CNBr-activated groups are transiently reactive; activity diminishes upon prolonged storage in aqueous media, especially at elevated temperature or high pH. Best performance when coupled soon after receipt and kept cold.
• Thermal/chemical tolerance: Agarose matrices generally tolerate 4–40 °C operationally and a wide pH range; however, reactive CNBr groups hydrolyze faster at high pH and temperature (use cool conditions during coupling). Exact allowable ranges are resin-specific and not specified for this item.

Important caveats:

• Do not dry the gel; drying collapses pores and irreversibly damages performance (general guidance).
• Avoid freezing (item-specific): Store at 2–8 °C, Do not freeze (from Product Data).

Quality and Grades

• Item-specific grade: BioReagent (from Product Data).

What BioReagent grade signifies (general explanation):

• Intended for research and laboratory use, suitable for biochemical workflows such as protein immobilization and affinity chromatography development.
• Typical expectations include low bioburden and consistent performance in common biochemical buffers. However, heavy-metal limits, endotoxin levels, and specific purity metrics are not provided here and can vary by lot; consult the CoA/Spec Sheet for details.

Matrix/reagent-specific quality considerations:

• Activation capacity: Practical coupling capacity (µmol of primary amine per mL of settled gel) is a key performance parameter for CNBr-activated agarose. Not specified for this item; refer to CoA/Spec Sheet.
• Bead characteristics: Bead size distribution, porosity, and crosslinking degree impact backpressure and binding of large biomolecules. Not specified for this item; refer to CoA/Spec Sheet.
• Leachables: Post-coupling washing protocols minimize leachable, unreacted groups and non-covalently bound ligand (general practice).
• UV background: For optical assays, low UV absorbance of the matrix and storage solution is advantageous; not specified for this item.

Stabilizers/preservatives:

• Any antimicrobial or storage preservative concentration is not specified for this item; refer to CoA/Spec Sheet. If present, remove or exchange by extensive washing or dialysis after coupling.

Compliance note:

• This product is labeled For research use only (from Product Data) and is not intended for diagnostic or therapeutic manufacturing without appropriate qualification.

Reaction and Applications

Focus: Covalent immobilization of ligands via primary amines to create custom affinity matrices (general/literature guidance expanded from the product name and typical use).

• Coupling chemistry: CNBr activation converts agarose hydroxyls into reactive cyanate esters/imidocarbonates. Primary amines on ligands (e.g., lysine ε-NH2 in proteins, aliphatic amines on small molecules) attack these groups to form stable isourea linkages.
• Use cases:
  • Preparation of antibody, enzyme, lectin, or small-molecule affinity columns for purification, depletion, or enzyme immobilization.
  • Immobilization of antigens/peptides for immunoprecipitation (IP), pull-down assays, or biomarker enrichment.
  • Creation of scavenger resins to remove contaminants or excess reagents from complex mixtures.
• Practical tips:
  • Desalt ligands into amine-free coupling buffer (e.g., 0.1 M NaHCO3, 0.5 M NaCl, pH ~8.3). Avoid Tris, glycine, ammonium salts during coupling.
  • Keep the resin cold (2–8 °C) prior to use; perform coupling at room temperature or 4–8 °C depending on ligand stability. Minimize coupling time for labile proteins.
  • After coupling, block remaining active groups with 1 M ethanolamine, pH ~8.0 (literature), then perform alternating high- and low-pH washes to remove adsorbed ligand.
  • Pack into gravity-flow columns or low-pressure systems; monitor backpressure if using pumps. Bead size/crosslinking (not specified for this item) determine flow limits.
• Performance notes:
  • Orientation: Random lysine coupling can reduce activity for some proteins; consider mild conditions, competing amines, or site-specific strategies if needed.
  • Regeneration: Many immobilized-ligand columns tolerate repeated cycles with appropriate clean-in-place; follow ligand-specific stability limits.

All numeric capacities, bead size, or pressure limits are not specified for this item; consult CoA/Spec Sheet.

Reaction Conditions

Typical coupling and post-coupling conditions for CNBr-activated agarose (general/literature guidance; adjust to your ligand):

• Pre-equilibration:
  • Wash resin with 1–3 column volumes (CV) of cold 1 mM HCl or deionized water to remove storage solution and stabilize reactive groups. Keep at 2–8 °C until coupling.
• Coupling buffer (amine coupling):
  • 0.1 M NaHCO3, 0.5 M NaCl, pH ~8.3 (amine-free). Alternative: 0.2 M NaHCO3/Na2CO3 pH 9.0 for less nucleophilic ligands.
• Ligand concentration:
  • Protein ligands: Often 1–10 mg/mL in coupling buffer; optimal value depends on desired capacity and protein stability. Not specified for this item; determine empirically.
• Temperature and time:
  • 20–25 °C for 1–4 h with gentle end-over-end mixing, or overnight at 4 °C for sensitive proteins. Activity decays over time—do not unduly delay coupling after washing.
• Blocking of residual active groups:
  • 1 M ethanolamine, pH ~8.0, 1–2 h at room temperature (gentle mixing).
• Post-coupling stringent wash:
  • Alternate 3–6 cycles of: (a) 0.1 M acetate, 0.5 M NaCl, pH ~4; and (b) 0.1 M Tris-HCl, 0.5 M NaCl, pH ~8–9 to remove adsorbed ligand.
• Column packing:
  • Pack at low pressure suitable for agarose beads; avoid high linear velocities that compress the bed. Pressure/flow limits are resin-specific and not specified for this item.
• Cleaning-in-place (CIP):
  • Depends on immobilized ligand stability. Common reagents include 0.5–1.0 M NaCl, brief low/high pH exposures, and up to 20% ethanol. Avoid conditions that denature or hydrolyze the ligand or the isourea linkage.

Always verify conditions at small scale before committing valuable ligands or samples.

Safety and Handling

Authoritative safety information must be taken from the product SDS. The following are general laboratory precautions for CNBr-activated agarose resins.

• GHS/Classification (item-specific): Not specified for this item; refer to SDS. No hazard pictograms or H-statements were provided in the Product Data.
• Primary hazards (general):
  • Irritation: Reactive surfaces and any storage preservatives can cause skin/eye irritation upon contact.
  • Cyanate/imidocarbonate reactivity: Activated groups can react with nucleophilic biomolecules; avoid contact with skin and eyes.
• PPE: Wear lab coat, safety glasses or face shield, and appropriate chemical-resistant gloves (e.g., nitrile). Use in a well-ventilated area or fume hood when preparing coupling buffers or handling concentrates.
• Handling tips (general):
  • Keep cold (2–8 °C) to minimize hydrolysis of reactive groups.
  • Pre-equilibrate with coupling buffer just before use; minimize delays between washing and ligand addition.
  • Avoid strong bases and high temperatures during activation/coupling to preserve activity.
• Incompatibilities (general): Prolonged exposure to high pH, strong nucleophiles without intended ligand, primary amine–containing blocking buffers before coupling, and oxidizing sterilants not validated for agarose can degrade the resin.
• First aid (overview):
  • Skin/eye contact: Rinse with water for 15 minutes; remove contaminated clothing; seek medical advice.
  • Inhalation of aerosols/sprays: Move to fresh air; seek medical attention if symptoms persist.
  • Ingestion: Rinse mouth; do not induce vomiting; seek medical attention.
• Waste: Dispose of used resin and buffers according to institutional and local regulations; treat as chemical waste containing organic amines if applicable.

Solvent Selection

This product is a solid-phase, aqueous gel matrix rather than a solvent. Traditional “solvent selection” does not apply. Instead, buffer compatibility and miscibility considerations relevant to coupling and chromatography are summarized below (general/literature guidance):

• Medium: Use aqueous buffers. The beads are insoluble but fully hydrated in water-based media.
• Coupling buffer (amine coupling to CNBr groups):
  • Typical composition (literature): 0.1 M NaHCO3, 0.5 M NaCl, pH ~8.3. High ionic strength suppresses nonspecific ionic interactions during coupling.
  • Avoid primary amine buffers (e.g., Tris, glycine) during coupling—they compete with the intended ligand.
• Washing/elution buffers:
  • Post-coupling alternating pH washes (e.g., acetic acid/sodium acetate pH ~4 and Tris/NaCl pH ~8–9) help remove non-covalently adsorbed ligand (literature practice).
  • Operational buffers for affinity use depend on the immobilized ligand and target.
• Additives tolerance (matrix-general): Agarose resins commonly tolerate salts, nonionic detergents, and common denaturants (urea, guanidinium) within moderate ranges; exact tolerances are resin-specific and not specified for this item.
• Organic solvents: Prolonged exposure to strong water-miscible organics (e.g., >20–30% v/v) may dehydrate or shrink the gel and reduce performance; brief exposure during cleaning-in-place can be acceptable depending on resin specification (not specified for this item).

When to choose CNBr-activated agarose versus alternatives:

• Choose CNBr activation for rapid, amine-directed immobilization without requiring carbodiimide chemistry, particularly for proteins with accessible lysines and when maintaining near-neutral to mildly basic pH is desired.

Storage and Reconstitution

Item-specific instructions (from Product Data):

• Storage Conditions: Store at 2–8 °C. Do not freeze.
• Shipped In: Wet ice.

Additional handling guidance (general/literature):

• Container handling: Keep tightly capped to prevent microbial contamination and evaporation. Avoid prolonged exposure to ambient temperatures; return to 2–8 °C promptly.
• Do not dry: Never allow the gel to dry; drying irreversibly damages pore structure and performance.
• Before first use ("reconstitution" concept for slurries):
  • Gently resuspend the slurry by inversion. If a preservative is present (not specified for this item), decant/storage solution and wash with 3–5 resin bed volumes of cold deionized water or 1 mM HCl, followed by coupling buffer, immediately before coupling.
  • Minimize time between washing and ligand addition to preserve reactive CNBr groups.
• Post-coupling storage: Store the immobilized-ligand resin at 2–8 °C in an appropriate buffer containing a compatible antimicrobial (e.g., 20% ethanol) if allowed by the ligand and institutional policies. If sodium azide is considered, ensure compatibility with downstream applications and safety protocols. Specific preservatives are not specified for this item; refer to CoA/Spec Sheet.
• Freeze–thaw: Avoid freezing at all times; ice crystal formation fractures beads.
• Shelf life/expiration: Not specified for this item; refer to CoA/Spec Sheet. As a general rule, activation decays with time and temperature; couple ligands soon after receipt for best performance.

For research use only (from Product Data).

Structure and Identity

Brief description: CNBr-activated agarose is a porous, crosslinked agarose bead matrix whose surface hydroxyls have been converted (via cyanogen bromide activation) to reactive groups that couple primarily with ligand primary amines to form isourea linkages.

• Item-specific identifiers (from Product Data):
  • SKU: C1523223
  • Product Name: CNBr-activated Agrose (cyanogen bromide–activated agarose)
  • CAS: Not specified for this item; refer to CoA/Spec Sheet.
  • CID: Not specified for this item; refer to CoA/Spec Sheet.
  • InChIKey: Not applicable for an insoluble, crosslinked polymeric matrix.
  • SMILES: Not applicable for an insoluble, crosslinked polymeric matrix.
  • Molecular Formula: Not specified for this item; refer to CoA/Spec Sheet. (Agarose is a polysaccharide polymer of alternating agarobiose units.)
  • Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet. (Beads are macroscopic particles with effectively polydisperse, very high MW.)

Structural features (general/literature):

• Backbone: Neutral polysaccharide composed of repeating agarobiose disaccharide units (β-D-galactopyranose and 3,6-anhydro-α-L-galactopyranose).
• Functionalization: Surface hydroxyls converted by CNBr activation to cyanate esters and imidocarbonate-type groups that rapidly react with nucleophiles, especially primary aliphatic amines on proteins/ligands.
• Morphology: Spherical, highly hydrated gel beads; pore size and bead diameter depend on the base agarose and crosslinking (not specified for this item).
• Stereochemistry: Multiple stereocenters inherent to the saccharide units; polymer is not a single defined stereoisomeric molecule.

2D description in words (general): Repeating sugar rings linked by glycosidic bonds, with pendant activated carbonate/imidocarbonate functions accessible on the bead surface for covalent ligand attachment.

Synthetic Utility

Although not a small-molecule reagent, CNBr-activated agarose is a powerful tool for synthetic biochemistry and chemical biology (general/literature):

• Functional group capture: The resin’s activated groups selectively react with primary amines to form isourea bonds, enabling immobilization of:
  • Proteins (via lysine residues), peptides (N-terminus or lysine side chains), amine-terminated oligonucleotides, or small molecules bearing aliphatic amines.
• Retrosynthetic perspective: By transferring a challenging separation into the solid phase, one can design syntheses where purification is accomplished by binding/elution rather than traditional chromatography (e.g., immobilized catalysts or affinity tags on intermediates).
• Immobilized catalysts: Enzymes or organocatalysts with amine handles can be tethered to the matrix to enable flow or batch catalysis with facile separation and reuse. Reaction engineering must consider mass transfer within the gel and solvent compatibility (aqueous preferred for agarose).
• Affinity capture in analytical workflows: Creating custom affinity sorbents for target enrichment prior to MS or HPLC analyses reduces matrix interference and improves sensitivity.
• Derivatization strategies:
  • Spacer arms: Coupling a short diamine or amino-PEG linker (via one end) can distance bulky ligands from the surface to improve accessibility; then cap remaining actives and couple the ligand via a secondary chemistry.
  • Site-selective immobilization: Protect or pre-block certain amines on the ligand to favor N-terminus coupling, or employ transamination/oxidation to create unique handles.

All resin-specific performance metrics (capacity, bead size) are not specified for this item and should be verified experimentally.

Target Specificity

Not applicable. CNBr-activated agarose is a general-purpose activated matrix without an inherent biological target. Specificity arises only after the user immobilizes a ligand (e.g., antibody, enzyme substrate analog, inhibitor). No item-specific antigen, epitope, species reactivity, clone, or isotype data apply.