Dextrin Magnetic Beads for MBP-Tagged Protein Purification

Overview

Description

Dextrin Magnetic Beads for MBP-Tagged Protein Purification

Tested applications

Affinity Purification of recombinant Proteins tagged with maltose binding Protein (MBP)

Properties

Form

Magnetic Beads in Suspension (20% slurry), supplied in phosphate buffered saline (PBS) pH 7.4, with 0.02% sodium azide as preservatives.

Storage instruction

2 to 8°C, do not freeze

Beads Diameter

40 µm

Binding Capacity

5 to 10 mg MBP fusion protein per 1 mL settled beads (5mL 20% slurry). 

Ligand

Dextrin

Matrix

Magentic Beads

Applications

Protocols

Buffer preparation

Binding/Wash buffer: 20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, pH 7.4

Elution buffer: 10 mM maltose in binding buffer

Regeneration buffer: 0.5 M NaOH

 

Purification Protocol

 

1. Sample preparation 

Adjust the sample to the composition of the binding buffer. For example, dilute the sample with binding buffer or buffer exchange using a desalting column.

 

Pass the sample through a 0.22 µm or a 0.45 µm ?lter and/or centrifuge it immediately before applying it to the column. If the sample is too viscous, to prevent it from clogging the column dilute it with binding buffer, increase lysis treatment (sonication, homogenization), or add DNase/RNase to reduce the size of nucleic acid fragments.

 

2. Purification of Recombinant MBP-Fusion Protein 

1). Equilibrate the beads with at least 5 beads volumes of binding buffer. 

2). Add the pretreated sample. Resuspension and incubation for 20-30 min.

3). Wash with 5 to 10 beads volumes of binding buffer for 3-5 times.

4). Elute with 1-5 beads volumes of elution buffer. 

5). After elution, regenerate the beads by following the procedure (see below).

 

3. After puri?cation, the beads should be regenerated as follows:

1). Regenerate the beads with 3 beads volumes of distilled water followed by 3 beads volumes of 0.5 M NaOH and 3 beads volumes of distilled water. 

2). Re-equilibrate the beads with 5 beads volumes of binding buffer (20 mM Tris-HCl, 200 mM NaCl, 1 mM EDTA, pH 7.4), then store in the same volume of 1*PBS before starting the next puri?cation.