CNBr-activated Agrose 4FF - BioReagent,70% v/v; 90μm

Cat. No.: C1523222

GRADE & PURITY BioReagent, 70% v/v; 90μm

Synonyms

CNBr-activated Resin

Storage

Store at -20°C

Shipped In

Ice chest + Ice pads

Application

Protein purification

Panoramica

CNBr-activated Agrose 4FF is a pre-activated agarose medium with 4% highly cross-linked agarose as the matrix, featuring excellent physical and chemical stability. The medium enables simple, direct, rapid and mild coupling, and can directly conjugate proteins and other biological macromolecules containing primary amino groups without a coupling spacer arm. It also allows multi-point coupling, making ligands not easy to detach. Meanwhile, it has strong hydrophilicity, low non-specific adsorption, wide universality and application scope, fast flow rate, good repeatability and easy process scale-up. It is mainly used for the separation and purification of biological molecules such as proteins.

Aladdin  CNBr-activated Agrose 4FF is stored in acetone, with a volume ratio of gel to protective solution of 7:3. The product specification is based on the actual gel volume.

Parameter Specifications

Item	Specification
Matrix	4% Highly cross-linked agarose
Coupling functional group	-NH₂
Activated group	CNBr
Average particle size	90 μm
Coupling conditions	pH 8–10, room temperature (20–25℃) for 2–3 h or 4℃ overnight
Coupling capacity	13–26 mg α-chymotrypsinogen/mL medium
Recommended flow rate	<250 cm/h
Recommended pressure	<0.15 MPa (1.5 bar)
pH stability ①	3–11 (long-term); 2–11 (short-term )
Chemical stability	Stable in common aqueous buffers after ligand coupling; denaturants such as 8 M urea or 6 M guanidine hydrochloride can be used ②
Storage conditions	Acetone; below -15℃
Shelf life	1 year

Notes:

① Long-term refers to the pH range where the medium remains stable for a long time without adverse effects on its subsequent performance. Short-term refers to the pH range for medium regeneration, cleaning-in-place and disinfection based on experience.

② The stability after ligand coupling is premised on the ligand’s tolerance to changes in pH or chemical environment.

Instructions for Use

1. Coupling of Functional Ligands

1.1 Buffer Volume

Solution	Composition	Volume Ratio
Solution A	1 mM HCl, 0.5 M NaCl (pre-cooled, pH 2–3)	≥20× gel volume
Solution B	0.2 M NaHCO₃, 0.5 M NaCl, pH 8.3	≥10× gel volume
Solution C	0.1 M Tris-HCl, pH 8.0	≥5× gel volume
Solution D	0.05 M Tris-HCl, 0.5 M NaCl, pH 8–9	≥15× gel volume
Solution E	0.1 M Acetic acid-sodium acetate, 0.5 M NaCl, pH 3–4	≥15× gel volume
Solution F	0.02 M PB, 0.15 M NaCl, pH 7.0–7.6	≥10× gel volume

1.2 Dissolve dry powder protein ligand with Solution B; replace the buffer of solution-state protein ligand with Solution B.

1.3 Place an appropriate amount of affinity chromatography medium stored in acetone in a glass filter funnel, and wash with 15× gel volume of pre-cooled Solution A.

1.4 After washing, add the medium to the ligand solution, mix well, and react at room temperature for 2–4 h or at 4℃ overnight.

1.5 After coupling, wash with 5× gel volume of Solution B to remove excess uncoupled ligand.

1.6 Wash with 3× gel volume of Solution C, then continue to block remaining active sites of the medium with Solution C at room temperature for 2–4 h.

1.7 Wash alternately with Solution D and Solution E for 3–6 cycles, 3× gel volume each time.

1.8 Finally, wash with 10× gel volume of Solution F to complete ligand coupling.

Coupling Notes

Coupling buffer: Bicarbonate or borate buffer is commonly used. Do not use Tris or other amino-containing buffers as coupling buffer; pH 8–10 is recommended for high coupling efficiency. Low pH affects coupling activity, but pH 6 may be used to reduce steric hindrance or preserve ligand bioactivity.

Add 0.5 M NaCl to reduce protein-protein adsorption.

Temperature & time: 2–3 h at room temperature (25℃); overnight (10–16 h) at 4℃.

Ligand concentration: 5–10 mg protein per mL medium is recommended. Excess ligand causes high ligand density, steric hindrance, reduced binding capacity, difficult elution and increased non-specific adsorption.

2.Column Packing

2.1 Packing Buffer Preparation

Purified water, ultrasonic degassing for 15 min.

2.2 Chromatography Medium Preparation

Calculate required medium volume (compression factor ~1.15) and weigh. Replace buffer to packing buffer via vacuum filtration. Prepare ~75% gel suspension with packing buffer.

2.3 Chromatography Column Preparation

Inspect column for integrity and cleanliness. Install lower end cap, tighten O-ring, fix column vertically and verify with level gauge. Remove air bubbles from bottom sieve with packing buffer via syringe, then cap. Add ~2 cm height of packing buffer to column.

2.4 Column Packing (Example: 16 mm diameter, 15 cm bed height)

Mix gel suspension, pour slowly into column with glass rod; fill with packing buffer if needed. Connect regulator to system, purge air bubbles at set flow rate then pause. Insert regulator at 45° and seal. Open bottom cap, set flow rate to 60 cm/h until bed interface stabilizes, then increase to 400 cm/h for 45 min, mark interface. Pause system, cap bottom, disconnect top, loosen regulator seal, lower upper end cap to 2 mm below gel surface, retighten. Connect column to system for column efficiency test.

3.Column Efficiency Determination and Evaluation

Column efficiency should be tested immediately after packing. Column performance is usually evaluated by theoretical plates per meter (N/m) and peak asymmetry factor (As). Higher column efficiency indicates stronger separation ability. For accurate results, the sample volume is 1.0% of column volume, and linear flow rate is controlled at 15–30 cm/h. Load the sample close to the column inlet to minimize dilution.

Item	Acetone Method	NaCl Method
Sample	1.0% (v/v) aqueous acetone	0.8 M NaCl aqueous solution
Sample volume	1.0% column volume	1.0% column volume
Mobile phase	Pure water	0.4 M NaCl aqueous solution
Flow rate	30 cm/h	30 cm/h
Detector	UV-280 nm	Conductivity

Column Efficiency Calculation

Calculate N/m and As from the UV (or conductivity) curve using the following formulas:

N/m = N/L

N = 5.54(V_R/W_h)² 

As = b/a

Notes: L = column height; V_R = retention volume; W_h= peak width at half-height; a = first half-peak width at 10% peak height; b = second half-peak width at 10% peak height.As should be between 0.8 and 1.8. N/m > 3000.

4. Separation and Purification

4.1 Column Equilibration

Flush column with binding buffer for ≥3–5 CV until effluent pH and conductivity match binding buffer; zero UV detector.

4.2 Sample Loading

Load sample filtered through 0.2/0.45 μm membrane. Loading volume depends on medium capacity, target molecule concentration and conditions.

4.3 Column Washing

Wash with binding buffer for 3–5 CV until UV 280 returns to baseline (or use optimized washing conditions).

4.4 Elution

Elute with elution buffer; collect fractions when target UV peak appears.

4.5 Cleaning and Storage

Regeneration: Alternate high-pH (0.1 M Tris-HCl, 0.5 M NaCl, pH 8.5) and low-pH (0.1 M sodium acetate-acetic acid, 0.5 M NaCl, pH 4.5) buffers for 3–5 cycles (2–3 CV each). Equilibrate with binding buffer (3–5 CV), flush with pure water to zero conductivity, then flush with 20% ethanol (2–3 CV). Store at 2–8℃.

Conditions vary by ligand; all buffers must be 0.22 μm filtered. Sample buffer should match equilibration buffer.

5. Cleaning-In-Place (CIP)

CIP removes strongly bound, precipitated or denatured contaminants that impair performance, increase backpressure and reduce flow rate. Recommended every 1–5 cycles.

Precipitated/denatured contaminants: 0.1 M NaOH (4 CV, 1–2 h contact) → binding buffer (≥5 CV); or 6 M guanidine hydrochloride (2 CV) → immediate binding buffer wash (≥5 CV).

Hydrophobic impurities: 0.1–0.5% non-ionic detergent (2 CV) → binding buffer (≥5 CV); or 70% ethanol (3–4 CV) → immediate binding buffer wash (≥5 CV).

6. Sterilization

Minimize microbial contamination with 70% ethanol (if ligand-tolerant) for 12 h, then flush with sterile binding buffer (≥5 CV).

7. Storage

Unused medium: Store below -15℃, protected from light, dry, ventilated and sealed; short-term storage at 4℃ (light-proof).

Coupled medium/column: Store sealed in 20% ethanol or 2% benzyl alcohol. Replace storage solution every 3 months to prevent microbial growth.

8. Linear Scale-Up

Lab-scale purification can be linearly scaled to pilot or production scale with these guidelines:

Keep residence time constant for stable dynamic binding capacity.

Select column volume based on binding demand; adjust bed height if needed.

Determine column diameter by flow rate; recommended bed height: 3–15 cm.

Maintain uniform sample concentration and consistent elution conditions.

Specifications

Sinonimi

CNBr-activated Resin

Specifiche e purezza

BioReagent,70% v/v; 90μm

Stabilità e conservazione

Store below -15℃ long term (12 months).

Condizioni di conservazione di stoccaggio

Store at -20°C

Spedito in

Ice chest + Ice pads

Questo prodotto richiede spedizione a catena fredda. I servizi di terra e altri servizi economici non sono disponibili.

Grado

BioReagent

Product Line

Prodotti biologici

Application Protocols

The following are general literature protocols for CNBr-activated agarose resins. They are provided as guidance and are not item-specific specifications.

1. Ligand Coupling (amine-directed)

• Wash: Slurry resin; wash 3–5× with 5–10 bed volumes (BV) of cold 1 mM HCl. Decant between washes.
• Equilibrate: Transfer to cold 0.1 M NaHCO3/Na2CO3, pH 8.3–9.0, 0.5 M NaCl.
• Add ligand: Mix with ligand solution (avoid Tris/glycine). Final resin:solution ratio typically 1:1 to 1:5 (v/v). Rotate 1–2 h at 0–4°C (up to overnight as needed).
• Block: Add ethanolamine to 1 M, pH 8.0–8.5; incubate 1–2 h to quench residual actives.
• Wash stringently: Alternate pH/high-salt washes (e.g., acetate pH 4/0.5 M NaCl and Tris or phosphate pH 8–9/0.5 M NaCl) for 3–6 cycles.
• Equilibrate/storage: Place in desired buffer (often with 20% ethanol for storage). Keep cold.

2. Column Packing (Fast Flow agarose)

• Prepare slurry (~50% v/v) in packing buffer; degas.
• Pack at recommended linear velocity to achieve desired bed height; check asymmetry and reduced plate height. Adjust flow to minimize channeling. Specific flow/pressure limits are Not specified for this item.

3. Immunoprecipitation (batch)

• Incubate sample with antibody-coupled beads 1–2 h at 4°C; wash 3–5× with buffer; elute with low pH glycine (pH 2.5–3) or chaotropic buffer; neutralize immediately.

Always confirm compatibility with the immobilized ligand.

Biological Roles

This product is an engineered chromatographic support; it does not have intrinsic biological activity. However, the base matrix (agarose) originates from marine polysaccharides and has properties relevant to biomolecular separations.

• Agarose origin (literature): Derived from red algae; composed of repeating disaccharide units of D‑galactose and 3,6‑anhydro‑L‑galactose. Forms thermoreversible gels due to double-helix aggregation and bundle formation.
• Biological compatibility (literature): Chemically inert, low nonspecific binding, and hydrophilic—traits that preserve protein structure during affinity capture.
• Functionalization: CNBr activation confers electrophilic groups that selectively couple to biomolecule primary amines, enabling creation of affinity ligands with defined orientation when using spacer strategies.
• Applications in life science workflows (literature):
  • Proteomics sample prep (immunodepletion, pull-downs)
  • Enzyme immobilization for pathway studies and biocatalysis
  • Lectin/aptamer immobilization for glycomics and interactomics

No medical, diagnostic, or therapeutic uses are claimed or supported for this product. For research use only (per Product Data).

Buffer Applications

Not a buffering reagent. CNBr-activated Agarose 4FF is a solid support used with buffers during coupling and chromatography.

Practical buffer guidance (literature/general):

• Coupling buffer: 0.1 M NaHCO3/Na2CO3, pH 8.3–9.0, often with 0.5–1.0 M NaCl to reduce nonspecific ionic adsorption. Avoid primary amines (e.g., Tris, glycine) which compete for coupling.
• Washing/equilibration: Phosphate-buffered saline (PBS) or phosphate buffer (pH 7.0–7.4) post-coupling. Alternate high/low pH washes (e.g., 0.1 M acetate, pH 4 with 0.5 M NaCl / 0.1 M Tris or phosphate, pH 8–9 with 0.5 M NaCl) to remove noncovalently bound ligand.
• Blocking solution: 1 M ethanolamine, pH ~8.0–8.5 to quench residual actives after coupling.
• Storage buffer for coupled resin: Typically neutral buffer with antimicrobial (e.g., 20% ethanol). Exact formulations and preservatives for this item are Not specified; validate compatibility with immobilized ligand.

Note: Always confirm buffer compatibility with the target ligand’s stability profile. Item-specific buffer recommendations are Not specified for this item; refer to CoA/Spec Sheet.

Green Alternatives

Considering sustainability and operator safety, selection among activation chemistries and process solvents can reduce environmental impact.

• Matrix choice: Agarose is a renewable, seaweed-derived polysaccharide; as a base material it is generally favorable versus petroleum-derived polymers (literature).

• Activation chemistry comparison (literature/general):

  • CNBr-activated agarose: Enables fast aqueous coupling at low temperature; CNBr is highly toxic during manufacturing (performed by supplier). User-stage hazards are moderate but reactive groups hydrolyze, generating benign byproducts. Waste minimization via efficient coupling and blocking is advisable.
  • NHS-activated agarose: High-efficiency amide formation near neutral pH, but NHS esters are moisture sensitive and often require anhydrous handling and cold-chain logistics.
  • Epoxy-activated agarose: No highly toxic reagents at point of use; broader nucleophile scope; typically requires longer times or elevated temperature, increasing energy input.
  • CDI-activated supports: Urea byproducts; good amine coupling but often need dry organic solvents.

• Process solvent considerations (literature): Favor water-based buffers; minimize organic cosolvents. If ligand solubility demands organics, choose lower-toxicity options (ethanol, isopropanol) over chlorinated solvents, limit percentage, and keep processes cold/short.

• End-of-life: Regenerate and reuse immobilized resins when possible; segregate and neutralize spent buffers; follow institutional green-chemistry waste hierarchies.

This item’s specific environmental metrics (e.g., E-factor, LCA) are Not specified for this item.

Pharmaceutical Uses

This product is for research use only (per Product Data) and is not intended for clinical manufacturing. Nonetheless, CNBr-activated agarose matrices are conceptually related to supports used in bioprocess development.

General/formulation-related roles (literature, non-clinical):

• Custom affinity sorbents for process development: Immobilize capture ligands (e.g., protein A/G mimetics, peptides) in early-stage purification scouting.
• Immobilized enzymes in route scouting: Create biocatalyst columns for screening transformations relevant to API intermediate synthesis.
• Impurity scavenging: Tailored amine-bearing ligands can be immobilized to remove specific process impurities during development.

Regulatory/compendial status: No pharmacopeial monograph is implied for this activated resin. Any use in regulated processes would require supplier qualification, extractables/leachables assessment, and process validation. Item-specific regulatory certifications are Not specified for this item; consult CoA/quality documentation.

No therapeutic, diagnostic, or in vivo applications are claimed. For research use only.

Physical Properties

Item-specific (from Product Data)

• Appearance: Not specified for this item; refer to CoA/Spec Sheet.
• Molecular weight, formula: Not applicable to a crosslinked polymeric support; not specified for this item.

General/literature values for CNBr-activated Agarose 4FF (non-specification)

• Bead form: Spherical, macroporous hydrogel beads (literature).
• Typical bead size distribution: ~45–165 µm for 4FF resins (literature, vendor-typical range; not a specification of this item).
• Matrix concentration: ~4% agarose (“4FF”), yielding relatively large pores suitable for proteins and antibodies (literature).
• Porosity/flow: Designed for low backpressure and high flow (“Fast Flow”) in gravity or low-to-moderate pressure columns (literature).
• Density/settled bed: Particle and bulk densities vary with hydration; commonly supplied as an aqueous or alcoholic suspension (literature). Exact values: Not specified for this item.
• Solubility: Insoluble in water and common solvents; swells in aqueous media (literature).
• Stability: Reactive groups hydrolyze in water, especially at elevated pH/temperature; store cold and couple promptly after washing (literature).

Practical implications

• Use cold buffers (0–4°C) and limit exposure to high pH prior to coupling to preserve activity (literature guidance).
• Avoid drying; maintain hydration to retain bead integrity (literature).

Quality and Grades

Item-specific (from Product Data)

• Grade/Purity: BioReagent.

What BioReagent grade implies (general guidance)

• Intended for research and laboratory applications, including biochemical coupling and purification workflows.
• Emphasis on lot-to-lot consistency, low bioburden/low endotoxin trends (when relevant), and suitability for coupling of proteins/peptides and subsequent chromatography. Exact bioburden/endotoxin limits: Not specified for this item; refer to CoA/Spec Sheet.
• UV background, metal content, and extractables are typically controlled for chromatography supports, but item-specific numerical limits are Not specified for this item.

Activation integrity

• CNBr activation level (i.e., available reactive capacity, commonly expressed as µmol active groups per mL settled resin) critically affects coupling density and performance. Item-specific capacity: Not specified for this item; refer to CoA/Spec Sheet.

Practical notes

• On receipt, maintain the cold chain per shipping/storage instructions to preserve reactive functionality.
• Validate each lot by small-scale test coupling (e.g., with a standard amine ligand) to confirm capacity and backpressure behavior in your system.
• Documented specifications (moisture content, preservative, slurry concentration) are best obtained from the lot CoA.

Reaction and Applications

CNBr-activated Agarose 4FF is a pre-activated solid support for covalent immobilization of ligands bearing primary amines. The resulting isourea linkage provides rapid, aqueous coupling suitable for biochemical separations.

Key application families (literature/general):

• Affinity chromatography media preparation: Immobilize antibodies, antigens, lectins, enzymes, peptides, or small amine-bearing ligands to create custom affinity resins for purification and enrichment.
• Immunoprecipitation/Co-IP: Prepare antibody-conjugated beads for batch or column capture of target proteins/complexes.
• Enzyme immobilization: Create immobilized biocatalysts for flow or batch reactors; improved stability and reusability in process development.
• Depletion columns: Couple albumin/IgG binders for plasma/serum depletion prior to proteomics.

Mechanistic notes (literature):

• CNBr activation forms imidocarbonate groups that react fastest with aliphatic primary amines at pH ~8–9 to give isourea linkages. Competing hydrolysis yields inactive hydroxyls; thus, cold conditions and prompt processing are critical.

Practical tips (literature):

• Avoid amine-containing buffers (e.g., Tris) during coupling; use carbonate or phosphate instead. Include salt to reduce ionic adsorption.
• After coupling, block residual actives with 1 M ethanolamine, pH 8.0, then alternate high/low pH and high-salt washes to remove noncovalently bound ligand.
• 4FF (“Fast Flow”) matrices permit higher linear flow rates and lower backpressure vs. non-FF agarose, facilitating scale-up.

Note: Exact ligand loading capacity and performance metrics are lot-dependent and Not specified for this item; consult CoA.

Reaction Conditions

Typical coupling and post-coupling conditions for CNBr-activated Agarose 4FF (literature guidance; not item-specific specifications):

• Pre-wash: Suspend resin and wash 3–5× with cold 1 mM HCl or cold deionized water to remove storage solvent/preservatives while keeping temperature 0–4°C.
• Coupling buffer: 0.1 M NaHCO3/Na2CO3, pH 8.3–9.0, optionally 0.5–1.0 M NaCl to suppress ionic binding.
• Ligand concentration: Often 1–10 mg/mL for proteins; adjust to achieve desired loading. For small molecules, use molar excess of amine functionality.
• Temperature/time: 0–4°C to room temperature; 1–2 h typical for initial coupling, up to overnight if needed. Keep cold to reduce hydrolysis.
• Mixing mode: End-over-end rotation or gentle stirring to prevent bead damage and ensure mass transfer.
• Blocking: 1 M ethanolamine, pH 8.0–8.5, 1–2 h at room temperature or 4°C to quench remaining active sites.
• Stringency washes: Alternate pH and ionic strength (e.g., 0.1 M acetate, pH 4 with 0.5 M NaCl; then 0.1 M Tris or phosphate, pH 8–9 with 0.5 M NaCl) to remove noncovalently bound ligand.
• Equilibration: Desired operating buffer (e.g., PBS) prior to use.
• Flow characteristics: 4FF supports generally permit higher linear velocities (literature: up to a few hundred cm/h in appropriately packed columns); exact operating backpressures depend on system and are Not specified for this item.

Always validate on small scale to optimize loading and activity for the specific ligand.

Safety and Handling

Item-specific (from Product Data)

• GHS classification, signal word, pictograms, H-statements: Not specified for this item; refer to SDS.
• Storage: Store at −20°C. Shipped in ice chest with ice pads.

General safety considerations (literature/good laboratory practice)

• CNBr-activated agarose contains electrophilic imidocarbonate groups; handle with appropriate PPE: lab coat, nitrile gloves, and safety glasses or face shield when splashes are possible.
• Avoid inhalation of aerosols or dust (if dried material is handled). Work in a fume hood when preparing or discarding acidic/basic solutions used in activation/coupling.
• Incompatibilities: Strong nucleophiles (e.g., concentrated amines) will consume reactive sites; acids/bases accelerate hydrolysis—control pH and temperature. Avoid sodium azide contact with copper plumbing in waste lines.
• First aid (overview; refer to SDS for authoritative guidance):
  • Skin/eye contact: Rinse with plenty of water for ≥15 min; remove contaminated clothing; seek medical attention if irritation persists.
  • Ingestion/inhalation: Move to fresh air; rinse mouth; seek medical advice.
• Waste: Quench unreacted active groups (e.g., ethanolamine blocking) before disposal. Treat biological ligands or process streams per institutional biosafety rules.
• Spills: Absorb aqueous suspensions with inert material; collect for disposal. Beads are slippery—clean floors thoroughly.

Always consult the product-specific SDS and institutional EHS guidance before use.

Solvent Selection

This product is a hydrophilic, crosslinked agarose resin used in aqueous systems. Solvent choice focuses on preserving reactive groups during coupling and maintaining bead integrity.

• Polarity/miscibility (literature): Fully compatible with water and common biological buffers. Insoluble but swellable in aqueous media. Limited compatibility with strong organic solvents; prolonged exposure to high organic content can dehydrate or shrink beads.
• Coupling media (literature): Mildly basic aqueous buffers (e.g., 0.1 M NaHCO3/Na2CO3, pH 8.3–9.0) to promote nucleophilic attack by primary amines. Maintain low temperature (0–4°C) to limit hydrolysis of active groups.
• Washing/removal of storage solvent (literature): Pre-wash with cold 1 mM HCl or coupling buffer to remove alcohol/preservative, then equilibrate in coupling buffer.
• Additives (literature cautions): Avoid primary amines (e.g., Tris, glycine) in coupling buffers—they compete with ligand. Prefer phosphate, carbonate/bicarbonate, or HEPES (amine-free) for equilibration. Include 0.5–1 M NaCl to reduce nonspecific ionic interactions when appropriate.
• Organic cosolvents (literature): Up to modest percentages of ethanol, isopropanol, or DMF can sometimes aid poorly soluble ligands, but monitor for resin dehydration and faster hydrolysis; minimize exposure time and keep cold.

Comparison (general):

• CNBr-activated agarose: Fast, amine-selective coupling in aqueous media.
• NHS-activated supports: Higher coupling efficiency at neutral pH but more moisture sensitive.
• Epoxy supports: More stable pre-activation; broader nucleophile scope, but longer coupling times.

Storage and Reconstitution

Item-specific (from Product Data)

• Storage conditions: Store at −20°C.
• Shipping: Ice chest with ice pads.

General guidance for CNBr-activated agarose (literature; complementing, not contradicting item data)

• Cold-chain handling: Keep resin cold at all times to minimize hydrolysis of reactive groups. Avoid repeated freeze–thaw if stored as a hydrated slurry; maintain consistent low temperature.
• Upon receipt: Allow to equilibrate to handling temperature in a cold room or on ice to avoid condensation. If supplied in an alcoholic preservative, keep container tightly closed until use.
• Reconstitution/Preparation for coupling:
  • Resuspend and wash beads 3–5× with cold 1 mM HCl or cold water to remove storage solvent.
  • Equilibrate in coupling buffer (e.g., 0.1 M carbonate/bicarbonate, pH 8.3–9.0, optionally 0.5–1.0 M NaCl).
  • Proceed immediately to ligand coupling; limit delays to reduce loss of activity by hydrolysis.
• Post-coupling storage (coupled resin): Store in a neutral buffer compatible with the immobilized ligand, commonly with 0.02% sodium azide or 20% ethanol as preservative (ensure compatibility and regulatory allowances). Exact preservatives for this item are Not specified for this item.
• Shelf life/retention of activity: Lot- and handling-dependent; monitor performance with a small-scale binding test. Item-specific expiry: Not specified for this item; refer to CoA/Spec Sheet.

For authoritative safety and handling details, consult the SDS.

Structure and Identity

Item-specific (from Product Data)

• Product name: CNBr-activated Agarose 4FF (SKU: C1523222)
• Grade: BioReagent
• Storage: Store at −20°C; shipped in ice chest with ice pads
• CAS, InChIKey, SMILES, Molecular Formula, Molecular Weight: Not specified for this item; refer to CoA/Spec Sheet.

General description (literature/typical for this material class)

• Matrix: Crosslinked agarose (4% agarose, “Fast Flow” grade), a neutral polysaccharide of repeating agarobiose units (D‑galactose and 3,6‑anhydro‑L‑galactose) forming a 3D hydrogel bead.
• Activation chemistry: Cyanogen bromide (CNBr) activation generating reactive imidocarbonate groups on the agarose backbone. These groups couple primarily to nucleophiles (notably primary aliphatic amines) on ligands to form isourea-type linkages.
• Structural features: Macroporous spherical beads; surface decorated with electrophilic carbonyl-derived imidocarbonates; no inherent chirality or aromaticity in the base matrix.
• 2D/“verbal” structure: Long chains of β‑(1→4)‑D‑galactose and α‑(1→3)‑3,6‑anhydro‑L‑galactose polysaccharide crosslinked into beads; CNBr activation introduces O‑(cyanate/imidocarbonate) groups along hydroxyls of the polymer.

Notes

• CNBr-activated agarose is supplied uncoupled; end users immobilize their chosen ligand via primary amines under mildly basic aqueous conditions, then block unreacted sites.

Synthetic Utility

From a synthetic chemistry perspective, CNBr-activated Agarose 4FF is a solid-phase support enabling covalent immobilization of primary amine-containing molecules, creating heterogeneous reagents or catalysts.

Utility highlights (literature/general):

• Heterogeneous catalysis: Immobilize organocatalysts or metal-binding ligands (via amines) to generate recyclable catalysts/scavengers with easy separation and reduced metal contamination risk.
• Affinity tags/handles: Attach capture ligands (e.g., biotin analogs, peptide motifs) to build selective scavenging resins for purification of complex mixtures.
• Solid-phase synthesis aids: While not a classical SPS resin, its hydrophilicity suits aqueous-phase transformations, bioconjugations, and flow-through applications where mass transfer is critical.

Reactivity scope (literature):

• Best for primary aliphatic amines; secondary amines couple more slowly; thiols/phenols are poor under standard conditions. Resulting isourea linkages are stable under neutral conditions but can be sensitive to strong acids/bases.

Operational advantages:

• Fast coupling at 0–4°C in water; minimal organic solvent required.
• 4FF matrix supports higher flow rates and lower backpressure versus non-FF agarose, aiding scale-up in columns.

Item-specific loading capacity, bead size distribution, and mechanical limits are Not specified for this item; verify experimentally and consult CoA.

Target Specificity

Not applicable. This is an uncoupled activated resin without an inherent biological target. Specificity arises only after the user immobilizes a chosen ligand (e.g., antibody, enzyme substrate analog, aptamer). No antigen/epitope/clone/isotype information applies.