PROTEINDEX™ Ni-Penta™ Agarose 6 Fast Flow, Chemical Stable, 50 mL, 11-0228-050, Marvelgent biosciences (CAT#: STEM-C-3664-LGZ)

PROTEINDEX™ Chemical Resistant Ni-Penta™ Agarose 6FF is a novel metal ion affinity chromatography (IMAC) medium, prefilled with nickel ions, mainly used for purification of low-abundance proteins or challenging proteins, requiring EDTA, DTT, β-ME and other stabilizers 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. In particular, Ni-Penta™ Agarose 6FF is ideal for capturing secreted poly-His-tagged proteins directly from cell culture supernatants of eukaryotic cells, including CHO cells, insect High Five™ cells* and Sf9 cells.

Highly cross-linked 6% agarose as the basic matrix carrier provides excellent mechanical stability for automated and large-scale purification of histidine-tagged proteins.

Product Highlights

EDTA-resistant
high selectivity, high purity
Maintains high 6xHis affinity in various buffers
No charging, minimal nickel ion leakage
Requires less imidazole
Resistant to NaOH, allows CIP of 1 M NaOH
Versatile, ready-to-use convenient format

Size: 50 mL
Ligand: Nickel
Binding capacity: >10 mg 6XHis-tagged protein/mL medium
Metal ion capacity: > 50 µmol Ni2+ / ml medium
Matrix: Highly cross-linked 6% agarose resin supplied as 50% slurry
Particle size: 45 μm - 165 μm
Maximum pressure: 0.3 MPa (3 bar)
Storage temperature: 2 - 8°C
Storage buffer: 1X PBS containing 20% ethanol

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

Due to its dedicated ligand for nickel ion immobilization, Ni-Penta™ resin is very tolerant to EDTA and its affinity for His-tag can be retained up to 10 mM in the presence of EDTA. To purify secreted proteins produced by eukaryotic cell cultures (such as insect cells, CHO cells, 293 cells, etc.), it is usually necessary to remove reagents incompatible with Ni-NTA resin by dialysis before loading the material onto the column. 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.

Chromatography Columns, 1 Adjustable End, 10-30bar, 25mm ID, 310mm Length, 25ba, GB-25-310-1E, G-BIOSCIENCES (CAT#-STEM-C-3349-LGZ)
Chromatography Columns, 1 Adjustable End, 10-30bar, 25mm ID, 460mm Length, 25ba, GB-25-460-1E, G-BIOSCIENCES (CAT#-STEM-C-3350-LGZ)
Chromatography Columns, 1 Adjustable End, 10-30bar, 25mm ID, 920mm Length, 25ba, GB-25-920-1E, G-BIOSCIENCES (CAT#-STEM-C-3351-LGZ)
Chromatography Columns, 1 Adjustable End, 10-30bar, 37mm ID, 100mm Length, 15 bar, GB-37-100-1E, G-BIOSCIENCES (CAT#-STEM-C-3352-LGZ)
Chromatography Columns, 1 Adjustable End, 10-30bar, 37mm ID, 230mm Length, 15 bar, GB-37-230-1E, G-BIOSCIENCES (CAT#-STEM-C-3353-LGZ)
Chromatography Columns, 1 Adjustable End, 10-30bar, 37mm ID, 310mm Length, 15 bar, GB-37-310-1E, G-BIOSCIENCES (CAT#-STEM-C-3354-LGZ)