Ni-charged MagBeads - 5-20mg 6xHis-tagged protein (27 kD)

Art No | L00295

Ni-charged MagBeads - 5-20mg 6xHis-tagged protein (27 kD)

130.00 CHF

Brochure

Size: 8mL (25% slurry)

The GenScript Ni-Charged MagBeads are developed for quick and efficient small-scale purification of polyhistidine-tagged proteins under native or denaturing conditions.

Description

The GenScript Ni-Charged MagBeads are average 40 μm in size, superparamagnetic beads with strong metal-chelating agent covalently bound to their surfaces.

They are pre-charged with nickel and ready to use for quick and small-scale purification of polyhistidine-tagged proteins. The beads are supplied as 25% slurry in phosphate buffered saline (PBS), pH 7.4, containing 20% ethanol. The Ni-Charged MagBeads have a binding capacity of 40 mg 6xHis-tagged protein per 1 ml settled beads (e.g. 4 ml 25% slurry).

Key Features

Quick and convenient separation accomplished by magnetic forces
High binding capacity with 40 mg of 6xHis-tagged protein/ml settled beads
Compatible with various reagents needed in the purification process (see table 1)
Can be regenerated for multiple uses
Extremely low nonspecific binding
Can be reused > 30 cycles*

Application

Purification of His-tagged proteins

Table 1 - Reagents Compatible with Ni-charged Magnetic Beads

Denaturants	Detergents	Reducing agents	Salts	Others
6 M Gu•HCl	2% Triton X-100	20 mM β-ME	4 M MgCl₂	50% glycerol
8 M Urea	2% Tween 20	1 mM DTT	5 mM CaCl₂	20% ethanol
	1% CHAPS		2 M NaCl	1 mM EDTA

*CIP was performed after each cycle, by incubating with 5 CV 0.1 M NaOH for 1 hr. Then magnetic beads were rinsed with excessive dd water and re-equilibrated with binding buffer before next purification cycle.

Storage: store at 4°C - do not freeze

Product manual (PDF)

Magnetic Beads Selection Guide

6xHistidine and Glutathione S-transferase (GST) are commonly used protein/peptide tags added to the end of a recombinant protein to aid in their purification or detection.

GenScript Ni-Charged MagBeads and Glutathione MagBeads provide simple, rapid and reliable methods for micro-scale purification of His- and GST-tagged fusion proteins from crude cell lysate prepared from bacteria, yeast or mammalian cell culture.

The Ni-Charged MagBeads are pre-charged with nickel and ready to use for micro-scale purification of 6xHis-tagged proteins under native or denaturing conditions. The Glutathione MagBeads are immobilized with Glutathione mainly through its central carbon on the beads surface for efficient GST binding.

All the protocols are easily performed by applying the magnetic separation technology. They do not require the use of columns or centrifugation and all the steps take place in a single tube. The MagBeads can be used in very small volumes to acquire high concentrated target proteins. All the advantages make Ni-Charged MagBeads and Glutathione MagBeads the ideal tools for micro-scale purification of His- and GST- tagged fusion protein.

Principle
The sample containing antibody is added to the Protein A MagBeads. The antibody will bind to beads during a short incubation. Then the beads‐bound antibody can be eluted off from the beads using a magnetic separation rack.

Magnetic separation eliminates the changes of micro tubes, minimizes the loss of sample and removes excessive steps of traditional centrifugation method.

For Expression screening

MagBeads Advantages:

Simpliﬁed handling of samples without the need for a chromatographic system
Ability to analyze several samples simultaneously thus reducing the total time required for analysis
Minimum sample loss and higher recovery than protein A resin

Case study 1:

For 5 mL volume, AmMag protein A magnetic beads compared with prepacked resin column

MagBeads: 500 μL AmMag protein A magnetic beads

Prepacked column: 1 mL prepacked Monoﬁnity A resin (combined with AKTA)

Samples containing human IgG1 produced in CHO cells which were continuously taken from a bioreactor during a two-week cell culture period.

Case study 2:

For 50 mL volume, AmMag protein A magnetic beads compared with pre-packed resin column

MagBeads: 500 μL AmMag protein A magnetic beads

Prepacked column: 0.6 mL Robocolumn A Resin (combined with Tecan)

Samples containing human IgG1 produced in CHO cells which were continuously taken out from day 14 cell culture period.

Time: Using MagBeads, 5 samples were puriﬁed in 60 minutes. Using column method, 5 samples were puriﬁed in 4 hours. MagBeads method saves 75% time.

Purity: Samples 1-5 were puriﬁed with MagBeads, sample 6 was puriﬁed by column puriﬁcation.

Using MagBeads method, 1 sample was puriﬁed in 1.5 hours. Using column method, 1 sample was puriﬁed in 3 hours.

MagBeads method saves 50% of time.

For large scale protein purification

AmMag protein A magnetic beads characteristics:

Simpliﬁed handling of samples without the need for a chromatographic system
No clarifying steps needed
Scalability: Simple capture of antibodies from large sample volumes (microliter to liliter scale)

Case study 3:

1 L antibody expression cell culture media (100 mg antibodies per L)

MagBeads: 5 mL AmMag protein A magnetic beads

Prepacked column: 5 mL Monofnity A Resin (combined with AKTA)

Samples containing human IgG1 produced in CHO cells which were continuously taken out from day 14 cell culture period

Supplier

Genscript
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Accessories

Art No | L00722

AmMag™ MR-mini magnetic rack for 2mL tubes

GenScript provides magnetic racks that help you purify proteins from small scale (≤ 2 mL), midi-scale (2 – 15 mL) and large-scale (15- 50 mL) cell cultures and lysates. These modular magnetic racks allow fast protein purification while simplifying the procedure.

230.00 CHF