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PROTEINDEX™ Ni-Penta™ Agarose, Chemical Stable, 100 mL, 11-0227-100, Marvelgent biosciences (CAT#: STEM-C-3671-LGZ)

Highlights

Direct purification of his-tagged proteins from eukaryotic cell culture supernatants

Due to its proprietary nickel ion-immobilizing ligand, Ni-Penta™ resin is very tolerant to EDTA and its affinity for his-tag can be maintained at 10 mM in the presence of EDTA. To purify secreted proteins produced by eukaryotic cell cultures (such as insect High Five™ cells* and Sf9 cells, CHO cells, 293 cells, etc.), it is often necessary to remove the Ni-NTA resin by dialysis before loading the material onto the column. Incompatible reagents. This is tedious, time-consuming and labor-intensive. With Ni-Penta resin, sample pretreatment can be skipped and media can be loaded directly onto the column for purification.

Cat Number: STEM-C-3671-LGZ

Application: For protein purification.

Model: 11-0227-100

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Description

Chemical Resistant Ni-Penta™ Agarose is a novel immobilized metal ion affinity chromatography (IMAC) medium prefilled with nickel ions that is primarily used for the purification of low-abundance or challenging proteins, in this case EDTA, DTT, β-ME and other stabilizers are required to maintain protein stability and integrity.

Nickel ions are very strongly bound to the matrix carrier, combined with the proprietary pentadate chelating ligand, and nickel ions are fixed through 5 sites to form a uniform chemically stable structure. This characteristic nickel ionic bond on Ni-Penta™ resins provides excellent resistance to EDTA, NaOH, β-ME and other reagents that are limited in purification protocols using traditional NTA and IDA functionalized resins.

Product Highlights

EDTA-resistant
high selectivity, high purity
Maintains high 6xHis affinity in various buffers
Ideal for capturing secreted poly-His-tagged proteins directly from eukaryotic cell culture supernatants
No charging, minimal nickel ion leakage
Requires less imidazole
NaOH-tolerant
Versatile, ready-to-use convenient format

Specification

Size: 100 mL
Ligand: Nickel
Binding capacity: >10 mg 6XHis-tagged protein/mL medium
Matrix: 4% agarose supplied as 50% slurry
Particle size: 45 μm - 165 μm
Maximum pressure: 0.1 MPa (1 bar)
Recommended flow rate: Gravity flow, <75 cm/hour
Storage temperature: 2 - 8°C
Storage buffer: 1X PBS containing 20% ethanol

Features

Direct purification of his-tagged proteins from eukaryotic cell culture supernatants

Due to its proprietary nickel ion-immobilizing ligand, Ni-Penta™ resin is very tolerant to EDTA and its affinity for his-tag can be maintained at 10 mM in the presence of EDTA. To purify secreted proteins produced by eukaryotic cell cultures (such as insect High Five™ cells* and Sf9 cells, CHO cells, 293 cells, etc.), it is often necessary to remove the Ni-NTA resin by dialysis before loading the material onto the column. Incompatible reagents. This is tedious, time-consuming and labor-intensive. With Ni-Penta resin, sample pretreatment can be skipped and media can be loaded directly onto the column for purification.

Purification of his-tagged proteins requires the presence of EDTA, DTT, β-ME and other reagents for maximum protein stability and integrity

PROTEINDEX™ Ni-Penta™ Agarose Resin is compatible with a wide range of buffers, including chelating agents (such as EDTA) and reducing agents (such as DTT), which are incompatible and should therefore be used in purification applications using traditional NTA or IDA resins. Avoid using. Ni-Penta™ resins help simplify sample preparation and allow purification of target proteins under their preferred conditions.

Reuse of columns that require cleaning-in-place with NaOH but no nickel charge

The stripping resistance of the Ni-Penta™ allows the column to be reused without charging the nickel ions, even after multiple CIP cycles with 1 M NaOH.

The amount of imidazole used in the elution buffer is limited due to the requirements of downstream applications or the sensitivity of the target protein to imidazole

Imidazole is often used to reduce non-specific binding to Ni-NTA resins. Ni-Penta™ Agarose Resin is highly selective for his-tags, so imidazole is not required throughout the purification to minimize non-specific binding. Imidazole is not required in the binding buffer, and 40 mM ~ 100 mM imidazole can effectively elute most of the target protein.

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