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P6611 Millipore

HIS-Select® Nickel Affinity Gel

(1:1 suspension in a 20% ethanol solution)

  •  NACRES NA.56

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Description

Application

HIS-Select®Nickel Affinity Gel has been used in the purification recombinant protein like EF-hand calcium-binding protein(S100A14), LIM homeobox transcription factor 1 alpha protein and sigma-1 receptor.

Nickel Affinity Gel used for the purification of histidine-tagged proteins. The unique, non-charged, hydrophilic linkage of the NTA chelate group to the agarose ensures high selectivity for histidine-tagged proteins and reduced non-specific binding of other proteins. Its high selectivity allows for one-step purification. Affinity gel can be used in either denaturing or non-denaturing conditions.

Features and Benefits

• High selectivity for higher purity
• Unique non-charged hydrophilic linkage reduced non-specific binding
• Binding capacity for histidine-tagged protein is greater than 15 mg/ml
• Binding under denaturing or non-denaturing conditions
• One-step purification

Linkage

It is also available with the EZview™ technology (Product Code E3528).

Physical form

1:1 suspension in a 20% ethanol solution

Legal Information

HIS-Select is a registered trademark of Sigma-Aldrich Co. LLC

Safety & Documentation

Safety Information

Symbol 
GHS02  GHS02
Signal word 
Warning
Hazard statements 
RIDADR 
UN1170 - class 3 - PG 3 - Ethanol, solution
WGK Germany 
3
Flash Point(F) 
89.6 °F
Flash Point(C) 
32 °C

Frequently Asked Questions

Can imidazole be used with HIS-Select® Nickel Affinity Gel, Product P6611?
For column chromatography, no more than 20 mM is suggested in the extract, equilibration, and wash buffers to prevent non-specific binding of proteins. No more than 250 mM is suggested for the elution buffers. Many proteins will elute with imidazole levels as low as 100 to 200 mM. For batch methods the imidazole concentration may have to be reduced or eliminated.When a protein is expressed at low levels, the presence of the imidazole limits the binding of the protein in the batch method but not when used in a column.
Can Tris buffers be used instead of phosphate buffer for HIS-Select® Nickel Affinity Gel, Product P6611?
Yes, Tris buffers should work.
Why won't my recombinant protein with a histidine-containing tag bind to HIS-Select® Nickel Affinity Gel, Product P6611?
Verify the pH and composition of sample and equilibration buffers. Make sure there are no chelating or reducing agents present in the extraction buffer. If using the batch mode, remove imidazole. Run the affinity purification under denaturing conditions. Run a Western blot of the extract to verify that the recombinant protein is present.
Can I use SDS with HIS-Select® Nickel Affinity Gel, Product P6611?
0.1% SDS has been used with some samples, with no adverse effects on the observed protein binding. However, SDS will effectively coat proteins and may block the binding to the column. It is probably very protein-specific and an SDS concentration that works for one protein may not work for another.
What needs to be done if the HIS-Select® Nickel Affinity Gel, Product P6611, resin turns brown on reuse?
During purification many protein extracts tend to discolor an affinity gel during the loading step. The original color will return after the wash or elution step. If the color is still not changing strip and recharge the affinity gel with nickel. Wash with EDTA and recharge with Nickel solution.
Which document(s) contains shelf-life or expiration date information for a given product?
If available for a given product, the recommended re-test date or the expiration date can be found on the Certificate of Analysis.
How do I get lot-specific information or a Certificate of Analysis?
The lot specific COA document can be found by entering the lot number above under the "Documents" section.
How do I find price and availability?
There are several ways to find pricing and availability for our products.Once you log onto our website, you will find the price and availability displayed on the product detail page.You can contact any of our Customer Sales and Service offices to receive a quote. USA customers: 1-800-325-3010 orview local office numbers.
What is the Department of Transportation shipping information for this product?
Transportation information can be found in Section 14 of the product's (M)SDS.To access the shipping information for this material, use the link on the product detail page for the product.
My question is not addressed here, how can I contact Technical Service for assistance?
Ask a Scientist here.
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