• USA Home
  • P6611 - HIS-Select® Nickel Affinity Gel

EMAIL THIS PAGE TO A FRIEND
P6611 Sigma

HIS-Select® Nickel Affinity Gel

Purchase

Description

Frequently Asked Questions

Frequently Asked Questions are available for this Product.

Application

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

50% suspension in 30% ethanol

Legal Information

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

Price and Availability


Amplified Detection
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.  That answer can also be found by clicking:  http://www.sigmaaldrich.com/sigma/bulletin/p6611bul.pdf
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.  The answer can also be found by clicking: http://www.sigmaaldrich.com/sigma/bulletin/p6611bul.pdf
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. These documents are located on the product detail page under Useful Links & Tools. Click on the following link to search for a Certificate of Analysis. Please click the following link to see the details on our Product Dating Information.
How do I get lot-specific information or a Certificate of Analysis?
A Certificate of Analysis is available by lot number and can be obtained through our Advanced Search Option: http://www.sigmaaldrich.com/catalog/AdvancedSearchPage.do
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 or view 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, or search here. 
My question is not addressed here, how can I contact Technical Service for assistance?
Use the option to the right to "Ask a Question" by email of a Technical Service Scientist.
Show more questions
Protocols & Articles
Peer-Reviewed Papers
15

References

Related Products

related product

Product #

Description

Add to Cart

08168 Timestrip Plus 8 °C

Technical Service:

Our team of scientists has experience in all areas of research including Life Science, Material Science, Chemical Synthesis, Chromatography, Analytical and many others.

Bulk Ordering & Pricing:

Need larger quantities for your development, manufacturing or research applications?