CNBr-activated Agrose (C008B-269624)

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.ParameterSpecificationMatrix4% AgaroseActivated groupCyanate esterAverage particle size90 μm (45–165 μm)Coupling conditionspH 8–10, room temperature (20–25 °C) for 2–4 h or 4 °C overnightCoupling capacity≥15 mg BSA/mL wet gelRecommended flow rate<75 cm/hWorking pH range3–11CIP pH range2–11Chemical stabilityStable in common aqueous buffers; compatible with denaturants such as 8 M urea or 6 M guanidine hydrochlorideStorage conditions100% acetone; 2–8 °CShelf life2 yearsInstructions for UseTo 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 PreparationBufferVolumeSolution A: 1 mM HCl, 0.5 M NaCl (pre-chilled, pH 2–3)≥20× medium volumeSolution B: 0.2 M NaHCO₃, 0.5 M NaCl, pH 8.3≥10× medium volumeSolution C: 0.1 M Tris, pH 8.0≥5× medium volumeSolution D: 0.05 M Tris, 0.5 M NaCl, pH 8–9≥15× medium volumeSolution E: 0.1 M acetate buffer, 0.5 M NaCl, pH 3–4≥15× medium volumeSolution F: 0.02 M PB, 0.15 M NaCl, pH 7.0–7.6≥10× medium volume2.Ligand PretreatmentTypically, 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 PretreatmentPlace 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 CouplingAfter 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.WashingAfter reaction, wash with 5× medium volume of Solution B to remove uncoupled protein ligand.6.BlockingWash 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 WashingAfter blocking, wash alternately with Solution D and Solution E for 3–6 cycles, using 3× medium volume per wash.8.StorageFinally, 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 Coupling1.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).RegenerationA 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.CleaningThe 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 Conditions2–8 °C, 100% acetone

Specifications & Purity: 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.ParameterSpecificationMatrix4% AgaroseActivated groupCyanate esterAverage particle size90 μm (45–165 μm)Coupling conditionspH 8–10, room temperature (20–25 °C) for 2–4 h or 4 °C overnightCoupling capacity≥15 mg BSA/mL wet gelRecommended flow rate<75 cm/hWorking pH range3–11CIP pH range2–11Chemical stabilityStable in common aqueous buffers; compatible with denaturants such as 8 M urea or 6 M guanidine hydrochlorideStorage conditions100% acetone; 2–8 °CShelf life2 yearsInstructions for UseTo 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 PreparationBufferVolumeSolution A: 1 mM HCl, 0.5 M NaCl (pre-chilled, pH 2–3)≥20× medium volumeSolution B: 0.2 M NaHCO₃, 0.5 M NaCl, pH 8.3≥10× medium volumeSolution C: 0.1 M Tris, pH 8.0≥5× medium volumeSolution D: 0.05 M Tris, 0.5 M NaCl, pH 8–9≥15× medium volumeSolution E: 0.1 M acetate buffer, 0.5 M NaCl, pH 3–4≥15× medium volumeSolution F: 0.02 M PB, 0.15 M NaCl, pH 7.0–7.6≥10× medium volume2.Ligand PretreatmentTypically, 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 PretreatmentPlace 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 CouplingAfter 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.WashingAfter reaction, wash with 5× medium volume of Solution B to remove uncoupled protein ligand.6.BlockingWash 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 WashingAfter blocking, wash alternately with Solution D and Solution E for 3–6 cycles, using 3× medium volume per wash.8.StorageFinally, 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 Coupling1.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).RegenerationA 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.CleaningThe 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 Conditions2–8 °C, 100% acetone

Grade: BioReagent

Storage Temperature: Store at 2-8°C;Do not freeze

Shipped in: Wet ice

UPC:

51111634

Weight:

1.23 Ounces

HazmatClass:

No

Quantity:

1

Unit of Measurement:

EA

WeightUOM:

LB

MPN:

C1523223-5ml

Country of Origin:

China

Product Size:

5ml

Harmonized Tariff Code:

2931.90.90