Zymo Research - His-Spin Protein Miniprep Kit

His-Spin Protein Miniprep™

HIGHLIGHTS

•	Fast 5 minute protocol to purify His-tagged proteins from cell-free extracts
•	Screen your recombinant colonies directly for protein production rather than for plasmid insert
•	Easy way to prepare pure protein for small-scale studies
•	Purified high-quality protein is suitable for enzyme kinetics, SDS-PAGE, MALDI-TOF, covalent modification, and other applications
•	No special instrumentation needed other than a benchtop microcentrifuge

DESCRIPTION

The His-Spin Protein Miniprep™ provides researchers with a fast His-tagged protein purification technology. The simplified procedure is based on our innovative protein purification chemistry and custom designed fast spin columns. Up to 1 mg of His tagged protein can be purified in 5 minutes and eluted in as little as 100 µl of His-Elution Buffer. The purified protein can be used directly for enzymatic assays, protein biochemical analyses, SDS-PAGE and other applications. The product has been optimized for maximal protein purity: a single protein band is visible by Coomassie blue staining on SDS-PAGE gel (Figure 1.). The straightforward spin – wash – elute protocol dramatically simplifies protein purification: get results in minutes, not hours.

ORDERING INFORMATION:

Products	Protocol	Cat No.	Size	Price
His-Spin Protein Miniprep™ Reagents Provided for 10 preps.	PDF	P2001	1 kit	$62.00
His-Spin Protein Miniprep™ Reagents Provided for 50 preps.	PDF	P2002	1 kit	$245.00

Components Available Separately
Zymo-Spin P1 Columns	P2003-1	50 pk	$37.00
Collection Tubes	C1001-50	50 pk	$5.00
	C1001-500	500 pk	$30.00
	C1001-1000	1,000 pk	$50.00
His-Affinity Gel	P2003-2	14 ml	$168.00
His-Binding Buffer	P2003-3	50 ml	$21.00
His-Wash Buffer	P2003-4	50 ml	$23.00
His-Elution Buffer	P2003-5	25 ml	$28.00

FREQUENTLY ASKED QUESTIONS:

How would you compare your His-Spin Protein Miniprep™ kit with similar products on the market?
Besides our His-Spin Protein Miniprep™, several other spin-format kits for purification of polyhistidine-tagged proteins are available on the market. They represent a huge advancement in protein purification techniques because of the speed and simplicity of use. Unlike our kit, however, most other products use metals (usually nickel or cobalt) immobilized on a dry, solid support. The protein to be purified actually comes into contact with the metal groups only after the beginning of centrifugation. Therefore, those protocols typically call for longer centrifugation times at low speeds to ensure enough time for the protein-metal interaction to occur. Because this requirement applies for the binding step as well as the washing and elution steps, the total purification time is typically longer (between 15-30 minutes) and substantially more inconvenient, than our His-Spin Protein Miniprep™.

The His-Spin Protein Miniprep™ utilizes nickel immobilized on agarose beads, which can be easily mixed with the proteins before centrifugation. This allows for immediate interaction of the proteins with the affinity gel. The centrifugation steps then serve only for separation of the filtrate from the gel. This way, we can take advantage of maximal centrifugation speeds and achieve as short as a 5 minute purification protocol. The shorter time is generally useful and may make all the difference if you are purifying a heat labile protein (many enzymes, for example) and do not have ready access to a refrigerated centrifuge. With other companies formats, spinning the sample several times for several minutes in an unrefrigerated centrifuge will expose the protein to higher temperatures, leading to decrease or even loss of activity. The Zymo's His-Spin Protein Miniprep™ is much more compatible with unrefrigerated centrifuges because 5 - 10 seconds spinning will have little to no impact on the temperature of the protein sample. It is also ideal for those simply wishing to save their valuable time.

REFERENCES:

Porath J, Carlsson J, Olsson I, Belfrage G. Metal chelate affinity chromatography, a new approach to protein fractionation. Nature. 1975 Dec 18;258(5536):598-9.

Hochuli E, Dobeli H, Schacher A. New metal chelate adsorbent selective for proteins and peptides containing neighbouring histidine residues. J Chromatogr. 1987 Dec 18;411:177-84.

Ueda EK, Gout PW, Morganti L. Current and prospective applications of metal ion-protein binding. J Chromatogr A. 2003 Feb 21;988(1):1-23.

Figure 1: E. coli cell extracts, containing indicated proteins expressed as a N-terminal hexahistidine fusion, as well as the proteins purified using His-Spin Protein Miniprep™, were analyzed by SDS-PAGE on a 15% gel, and stained with Coomassie Blue. The recombinant proteins were purposely expressed to a low level to demonstrate efficiency of the His-Spin Protein Miniprep™.