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Ni SepFast BG: Purification of a 6 x His tagged
hydrolase from unclarified E.coli. cell lysate in a batch operation
Lane 1: Molecular marker;
Lane 2: Unclarified cell lysate;
Lane 3: Cell lysate pass through;
Lane 4: Wash with binding buffer (20 mM phosphate, 20 mM imidazole and
500 mM NaCl, pH 7.4);
Lane 5: Wash with 50 mM imidazole in the binding buffer;
Lane 6 and 7: Eluates at 500 mM imidazole in the binding buffer, pH
7.4.
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Key benefits:
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Best performance : price ratio for batch or gravity flow purification
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Low product cost due to the novel manufacturing processes
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High binding capacity (could be over 40 mg/ml)
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Fast and easy protein purification from unclarified or clarified
samples
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All purification steps (binding, washing and elution etc) be done in a
single column (supplied) no matter if it is batch or gravity flow
operation
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Two gravity flow columns (BG-30 and BG-5) supplied for free
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No expensive chromatography system or column is required
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Choice of four pre-charged metal ions
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Very low metal leakage
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Full protein purification manual supplied
Its rough surface and carefully controlled pore structure allows fast
access of immobilised metal ligands by target protein molecules. Also,
its high mechanical strength permits liquid passing through gravity
columns at decent flow rates. Clarified or unclarified cell lysates
(for intracellular proteins) or culture broths (for extracellular
proteins) can be directly processed with IMAC SepFast BG. All these
translate into high process flexibility and higher protein yield at
shortened purification time.
To facilitate the purification process with as little hassle as
possible in batch or gravity flow operations, two specially designed
empty columns are supplied with the resins. The larger column BG-30
has a working volume of 30ml with volumetric marks in the first 20ml
(at 1ml interval). The smaller one BG-5 has a working volume of up to
5ml. There is a sintered porous mesh underneath each column to
withhold chromatography particles. Both lids in the bottom and the top
of the column are provided. Therefore, the whole protein purification
process, i.e. resin equilibration, binding, washing, elution and
regeneration (or clean-in-place) can be done within the same column in
a very economical way.
For further process flexibility, the desired
volume of resin can be loaded to the column of choice without any
restriction. In comparison, the pre-packed columns in marketplace have
fixed resin volumes (e.g. typically 0.1ml or 1 ml) that might not be
desirable. The smaller column is ideal for working resin volume of
0.1ml to 1ml. The larger one is ideal for resin volume of 1ml to 10ml. These columns can be reused after proper cleaning or used in a
disposable way. Extra columns can be ordered in separate.
IMAC SepFast BG or its four metal charged formats i.e. Ni SepFast BG,
Cu SepFast BG, Co SepFast BG and Zn SepFast BG, is highly stable and
compatible to a wide range of chemicals (e.g. detergents, denaturing
reagents and reducing reagents etc) commonly experienced in protein
purification processes, which means that more flexible operations can
be developed for the best performance.
A screening kit containing four types of immobilised metal ions is
available for scouting the best metal ion to given proteins.
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