High Density Aminoethyl Agarose Beads (4%)

High Density Aminoethyl Agarose Beads (4%) allows a covalent binding of agarose to carboxy group amino acids of the target ligand. This product is adequate to work in batch or column purifications (Low Pressure). The resin has a high amount of aldehydes groups (40-60 µmoles diaminoethyl /ml gel) providing a very good option to conjugate affinity ligands improving the stability of the ligand. The backbone of the resin is 4% agarose beads and the pore size is adequate to work with enzymes, proteins & antibodies.

Bead Geometry & Size	Spherical,Standard: ~50-150 µm
Crosslinked	Yes
Agarose %	4%
Matrix	Amino Groups
Activation Degree (μmol diaminoethyl/ml gel)	40-60
Antimicrobial Agent	20% Ethanol

Target	High Density Aminoethyl Agarose Beads (4%)
Storage	Store at 2-8°C.
Availability	Shipped within 5-10 working days.
Note	This product is for research use only.
Directions for use	Aminoethyl resins allow the covalent binding of agarose to ligands, which can be enzymes, proteins, peptides, antibodies or biomolecules. The amino groups of the resin react with exposed carboxyl groups of the ligand. The result of the biomolecule immobilization is a stable and reusable resin for affinity purification in batch, spin column or gravity procedures. The following procedure is suitable for the immobilization of ligands in batch or column procedures. Elimination of preservatives: Determine the quantity of Aminoethyl Resin needed for immobilization. The Resin is supplied as a 50% slurry in preservative. Note: 1 ml gel corresponds to 2.0 ml of the supplied suspension. Wash the Aminoethyl Agarose Beads with distilled water using a medium porosity sintered glass funnel (for batch immobilization) or a gravity column (for column immobilization). Batch immobilization: Manually shake the resin bottle to obtain a homogeneous suspension of beads and preservative. Invert the bottle several times and then filter the resin and put it in a container. 0.7 g of dried gel corresponds to approximately 1.0 ml of settled gel. Gravity column immobilization: Invert the resin bottle several times, and then pipette the desired volume into an empty gravity column. Note that the resin is supplied in an aqueous slurry containing preservative (50:50), so it is necessary to pipette double the volume of liquid to obtain the desired amount of gel. Sample preparation: Prepare the ligand solution and test the activity and/or absorbance at 280 nm. Prepare a solution of 8.85 ml distilled water, 0.19 g 1-(3-dymethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDC) and add the ligand. To find an appropriate concentration of ligand, albumin may be used as indicator, since it binds in similar proportions according to the table below. Aminoethyl (µmol/ml gel) Immobilized BSA (mg/ml gel) 3-6 ~5 15-25 ~14 40-60 ~30 If the ligand is not stable, run the remaining steps in a cold room. Coupling: Add 1 ml Aminoethyl Agarose Beads to the previous solution. Stir gently, take aliquots of the suspension and test the activity and/or absorbance at 280 nm. Continue gentle stirring for 1-3 hours or until the activity measurements remain constant, which indicates complete immobilization. Avoid magnetic stirring. Stirring should not exceed 3 hours as this can lead to decomposition, however, if this step is carried out in a cold room, the stirring time can be extended. Wash the suspension with distilled water to eliminate excess reagents, then with 1.0 M NaCl, and finally with distilled water. The ligand is now bound to the aminoethyl matrix and can be stored in a 0.03% sodium azide solution (4-10 °C).

Research Articles on High Density Aminoethyl Agarose Beads (4%)

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