Clontech Laboratories His60 Ni Superflow Cartridges Manual do utilizador - Página 4
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B. His60 Ni Superflow Resin
Clontech's His60 Ni Superflow Resin is a high-capacity Ni-IDA resin that has been optimized for the efficient
one-step purification of expressed his-tagged proteins from bacterial, yeast, mammalian, and baculovirus-infected
cells. The combination of the high density of nickel (II) ion and the high flow rates of the Superflow 6 agarose
beads allow the efficient capture of target his-tagged proteins. Up to 60 mg of his-tagged protein can be adsorbed
onto 1 ml of His60 Ni resin (data based on purification of AcGFP1).
The His60 Ni Superflow Cartridge Purification Kit (5 x 1 ml) provides 5 prepacked His60 Ni Superflow
Cartridges (each containing 1 ml of resin), as well as all the buffers needed for protein extraction and purification
(also available separately as the His60 Ni Buffer Set)—see Section II.
The His60 Ni Cartridge Purification Kit (5 x 5 ml) provides 5 prepacked His60 Ni Superflow Cartridges
(each containing 5 ml of resin), as well as all the buffers needed for protein extraction and purification (also
available separately as the His60 Ni Buffer Set)—see Section II.
Limitations of His60 Ni Superflow Resin
Please note the following recommendations when using His60 Ni Superflow Cartridges:
Do not use chelator-containing protease inhibitors or other additives, EDTA, or strong reducing agents
(see Table 2 and the note below regarding the use of reducing agents).
For automated liquid-chromatography (LC) applications, use highly pure, low-absorbance imidazole
(Fisher, Product No. BP 305-50). Always filter buffers through a 0.45 μm filter and degas before use.
His60 Ni allows protein purification under either native or denaturing conditions. The resin is compatible
with multiple denaturants and detergents (Table 2).
NOTE: Using βME as a reducing agent with His60 Ni Superflow Resin sharply reduces protein yield. If high
levels of βME are required for purification, we strongly recommend using TALON resin, which provides high
yields of the target protein in up to 30 mM βME.
PT5143-1
031716
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Clontech Laboratories, Inc. A Takara Bio Company
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