1. Overview

Magarose-DEAE is a weak anion-exchanged magnetic bead with fast magnetic responsiveness, abundant ion exchange capacity, and extremely high protein binding ability. The ion exchange ligand is diethylaminoethyl (DEAE), which can maintain a stable high protein binding capacity in the operating range of pH 3-12. Magarose-DEAE magnetic beads eliminate the need for pre-treatment of crude protein samples (e.g., tedious centrifugation, time-consuming and laborious filtration operations), flow rate and column pressure, and expensive chromatography equipment. For skilled operators, high-purity protein of interest can be extracted in a short time, and parallel processing of multiple samples can be easily achieved for high-throughput protein purification.

2. Product characteristics

Product Name	Magarose-DEAE
Item number	PMAG006
Magnetic bead concentration	25%(v/v)in 20% ethanol
Total ionic capacity	110~170 μmol/ml magnetic beads
Medium	6% cross-linked magnetic agarose
Particle size	20-45μm
Ligand binding	~110 mg BSA/ml beads
Save	Store at 2~8°C for one year

3. How to use

1. Product advantages

w Fast magnetic response speed, reducing operation time;

w The magnetic beads have good dispersion and resuspension, which improves the convenience of operation;

w The ligand has good physicochemical stability, which improves the reliability and reproducibility of experimental results.

2. Procedure (take purification of BSA-containing protein samples as an example).

1) Magnetic bead pretreatment (balance): Oscillate the magnetic bead vortex for 30 s, and fully resuspend it; A certain amount of 25% (v/v) magnetic bead suspension was placed in a 50 mL centrifuge tube. Magnetic separation, discard the supernatant, add 20 mL of equilibration buffer to wash the beads 3 times, and mix vertically for 2 min each time.

NOTE: To obtain the maximum recovery of the target protein, the experimenter needs to add excess magnetic beads, which are generally greater than 20% of the protein binding. For samples with lower abundance of target proteins, the recovery of the target protein from the magnetic beads will be reduced, so the amount of magnetic beads should continue to be increased.

2) Protein adsorption: Add the pretreated magnetic beads to the sample solution containing BSA protein, swirl for 30 s, and mix well in a vertical mixer for 30~60 min, so that the sample and the magnetic beads are fully contacted and adsorbed, and then magnetically separated and the supernatant is discarded.

Note: For more effective adsorption of binding substances, the equilibration buffer should contain a lower ionic strength, and the pH value should be at least one pH unit different from the isoelectric point of the target protein, and the pH of the selected salt buffer should fluctuate within 0.5 pH. The adsorption time of the target protein and magnetic beads is related to the properties of the protein itself.

3) Magnetic bead washing: add 20 mL of equilibrium buffer, resuspend the magnetic beads with vortex oscillation for 30 s, then magnetically separate, and discard the supernatant; This operation is repeated 3 times.

4) Protein elution: Add an appropriate amount of eluent to the centrifuge tube that has completed the magnetic bead washing mentioned above for protein elution. Quickly resuspend the magnetic beads, then place the centrifuge tube in the vertical mixer for 10~15 minutes to mix and then magnetically separate, and collect the supernatant into a new centrifuge tube. There are two main elution methods: high-salt elution (equilibration buffer containing 1-2 M NaCl) and low-pH elution (choose a pH range below the isoelectric point of the target protein).

5) Magnetic bead regeneration: Generally wash with 2 M NaCl solution 3~5 times, and then rebalance with equilibration buffer. After multiple uses, impurities such as precipitated protein, strong hydrophobic protein, lipoprotein and other impurities are non-specific adsorbed to the magnetic beads, and in order to ensure the efficiency of magnetic beads, it is recommended to clean in situ (CIP).

6) Cleaning in situ (CIP): wash the beads twice with 1.0 M NaOH, 70% ethanol or 30% isopropyl alcohol, and pure water; Finally, add 20% ethanol to resuspend the beads and store them at 2~8°C.

3. Precautions

1) This product should not be frozen, dried or centrifuged. Operations such as freezing, drying, and centrifugation can cause magnetic bead agglomeration, which is not easy to resuspend and disperse, and affects the chemical activity of functional groups on the surface of the magnetic beads.

2) Before using this product, be sure to keep the magnetic beads in a uniform suspension state.

3) During use, the magnetic beads can be washed 2~3 times with purified water or buffer to remove the ethanol in the preservation solution.

4) This product needs to be used with magnetic separation equipment.

5) Salt concentration and pH value will affect the binding and elution of specific proteins, and customers need to explore the binding and elution conditions of different proteins to ensure the purification amount and purity of proteins.

This product is for research use only!