MagListo™ His-tagged Protein Purification Kit (5 reactions)

MagListo™ His-tagged Protein Purification Kit allows easy purification of His-tagged proteins through magnetic separation by Ni-NTA Magnetic Nanobeads and buffers. With an average diameter of 400 nm, the beads have large surface areas with excellent binding capacity and can be recovered quickly without any loss through their strong magnetic force. These characteristics can quickly reduce the experimental time and the increase the yield. About 3~4 mg of 6x His-tagged proteins can be bound on to about 1 ml of Magnetic Nanobeads.

※This product is shipped with ice pack.

$270.00

Cat. No.

K-7200

Product Overview

Features and Benefits

Quick isolation
His-tagged proteins can be purified quickly and easily.
High-purity
Spherical shape magentic beads with good binding ability allow high-purity isolation of target proteins
Efficient screening
Use of crude cell lysate for efficient screening
Wide Application
Broad compatibility with all recombinant His-tagged protein purification with His-tag expressed in vivo (E. coli expression) or in vitro (cell-free expression).

Technical/Specs

▶ Components

Components	Pack Size	Storage Temperature
Ni-NTA magnetic silica resin	5 X 1 mL (10% slurry, v/v)	4°C
Binding/Washing buffer	100 mL
Elution buffer	15 mL
Neodymium magnet	3 ea
User's Guide	1 ea

▶ Procedure

Cell-shredded samples can be prepared in advance and processed according to the manual of this kit to purify 6x His-tagged protein.

※ Note
1. The neodymium magnet included in this kit can be used for purification.
2. This kit can be purified using centrifuge without neodymium magnet.
3. Bioneer's MagListo™-2(TM-1010), magnetic particle purification system, can make it easier and faster to purify 8 samples at a time.

▶Experiment Results

1. Protein purification using MagListo™ His-tagged Protein Purification Kit

Figure 1. Purification of 6x His-tagged protein.
- M: Protein broad marker (Bioneer, D-2010)
- L: Loading sample
- 1: Unbound sample
- 2: Eluted sample

2. Protein purification of various sizes from 25 kDa to 94 kDa using MagListo™ His-tagged Protein Purification Kit

Figure 2. Protein size effect on purification.
- M : Protein broad marker (Bioneer, D-2010)
- 1 : Size 94 kDa his-tagged protein
- 2 : Size 67 kDa his-tagged protein
- 3 : Size 38 kDa his-tagged protein
- 4 : Size 27 kDa his-tagged protein
- 5 : Size 25 kDa his-tagged protein

3. Comparison with other companies' magnetic bead products

Figure 3. Comparison with other products.
- M : Protein broad marker (Bioneer, D-2010)
- L : Loading whole sample
- 1 : MagListo™ His-tagged Protein Purification Kit (K-7200)
- 2 : Company A
- 3 : Company B
- 4 : Company C
- 5 : Company D

Literature/Support

◈ Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001 – certificate

• EN ISO 13485 - certificate

Ordering Info

Cat. No.	Product Description	Option
K-7200	MagListo™ His-tagged Protein Purification Kit	1 ml x 5

FAQ

Low protein yield

Expression of soluble protein is poor.
Optimize the expression level of the soluble target protein by adjusting temperature of fermentation, induction time and concentration of inducer.
Protein forms inclusion body.
If the protein forms inclusion bodies, it is unable to bind to the Ni-NTA magnetic beads. Optimize the expression process to minimize protein aggregation. Or purify aggregated protein under denaturing condition.
6x His-tag does not bind to the magnetic beads.
If the structure of 6x His-tag is partially hidden, it is unable to bind to the Ni-NTA magnetic beads. Move the tag to the opposite end of protein. Or purify the protein under denaturing conditions. If the binding conditions are incorrect, optimize both pH value and imidazole concentration of the binding buffer. The binding buffer provided in the kit contains 10 mM of Imidazole, and its pH is 8.0. For a more detailed information, refer to the manual.
Target protein does not elute in elution process.
Elution buffer conditions are too mild to elute protein. Increase imidazole concentration or decrease pH value. The elution buffer provided in the kit contains 500 mM of Imidazole, and its pH is 8.0. For a more detailed information, refer to the manual.

Poor protein purity

Buffer conditions are incorrect.
Optimize pH value and increase imidazole concentration of binding/washing buffer to avoid unspecific interaction. The binding/washing buffer provided in the kit contains 10 mM of Imidazole, and its pH is 8.0. For a more detailed information, refer to the manual.
Insufficient washing step.
Increase volume of washing buffer.
Column is too large.
Reduce the amount of Ni-NTA resin to reduce unspecific interaction. The Ni-NTA magnetic nanobead solution provided in the kit contains 100 mg of beads per 1 ml.

Protein is eluted during binding and washing step

Buffer conditions are incorrect.
Lower the concentration of imidazole or increase the pH value slightly. The washing buffer provided in the kit contains 10 mM of Imidazole, and its pH is 8.0. For a more detailed information, refer to the manual.
6x His-tag weakly bind to the magnetic beads.
If the structure of 6x His-tag is partially hidden, it might detach from Ni-NTA beads during the washing step. Reduce stringency of the washing buffer. Or purify under denaturing conditions. The washing buffer provided in the kit contains 10 mM of Imidazole. For a more detailed information, refer to the manual.

Selection Guide

The BIONEER products for recombinant protein synthesis and purification are categorized as the following, Those can be selected according to the purpose.

To apply for the ExiProgen™ demo, plase contact sales@bioneer.co.kr .

Process

Product type

Equipment used

Service

DNA Template

Preparation