HIS-Select® Nickel Affinity Gel

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


Quality Level


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




pkg of 1 mL
pkg of 100 mL
pkg of 25 mL
pkg of 5 mL
pkg of 500 mL


1.5-2.4 mL/mL (suspension in packed gel)


protein purification: suitable


faint blue to very dark blue


6% Beaded Agarose


>15 mg/mL, gel binding capacity (protein)(with an approx. 30 kDa protein)

transition temp

flash point 32 °C (closed cup)

storage temp.


General description

HIS-Select® Nickel Affinity Gel is an immobilized metal ion affinity chromatography (IMAC) product, used for the purification of His-tagged proteins. While the unique, non-charged, hydrophilic linkage of the proprietary quadridentate NTA chelate group to the beaded agarose charged with nickel ensures high selectivity for small to medium scale His-tag protein purification, it also results in reduced non-specific binding of other proteins. HIS-Select Nickel Affinity Gel is selective for recombinant proteins with His-tags and exhibits low non-specific binding of other proteins. The selectivity can be modulated with the inclusion of imidazole during chromatography.
HIS-Select® Nickel Affinity Gel is durable and can capture the recombinant proteins with histidine tags at a high flow rate. Recombinant proteins with histidine tags are bound using either native or denaturing conditions.


HIS-Select® Nickel Affinity Gel has been used in the purification of recombinant proteins like EF-hand calcium-binding protein (S100A14), LIM homeobox transcription factor 1 alpha protein, sigma-1 receptor as well as harpin, stable protein 1 (SP1), and BCR-ABL fusion protein.

Features and Benefits

  • High selectivity for higher purity.
  • Unique non-charged hydrophilic linkage reduces 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.
  • Minimizes unwanted ionic interactions.
  • Minimal nickel leaching.
  • Bead size: 45-165 μm.


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

Physical form

1:1 suspension in a 20% ethanol solution

Storage and Stability

HIS-Select Nickel Affinity Gel is stable for at least one year when stored properly. The HIS-Select Nickel Affinity Gel should be cleaned after each use and an antimicrobial agent such as 20% ethanol should be added to the storage buffer.

Legal Information

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



Signal Word


Hazard Statements


UN1170 - class 3 - PG 3 - Ethanol, solution

WGK Germany


Flash Point(F)

89.6 °F - closed cup

Flash Point(C)

32 °C - closed cup

  1. 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.

  2. Can Tris buffers be used instead of phosphate buffer for HIS-Select® Nickel Affinity Gel, Product P6611?

    Yes, Tris buffers should work.

  3. 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.

  4. 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.

  5. 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.

  6. 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.

  7. 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.

  8. 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.

  9. 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. 

  10. My question is not addressed here, how can I contact Technical Service for assistance?

    Ask a Scientist here.

Purification and characterization of the guinea pig sigma-1 receptor functionally expressed in Escherichia coli
Ramachandran S, et al.
Protein Expression and Purification, 51(2), 283-292 (2007)
S100A14, a member of the EF-hand calcium-binding proteins, is overexpressed in breast cancer and acts as a modulator of HER2 signaling
Xu C, et al.
The Journal of Biological Chemistry, 289(2), 827-837 (2014)
F Weerkamp et al.
Leukemia, 23(6), 1106-1117 (2009-04-24)
BCR-ABL fusion proteins show increased signaling through their ABL tyrosine kinase domain, which can be blocked by specific inhibitors, thereby providing effective treatment. This makes detection of BCR-ABL aberrations of utmost importance for diagnosis, classification and treatment of leukemia patients....
Leucine zipper-like motifs of HrpZPss are not essential to induce hypersensitive response in tobacco
Anil K, et al.
Journal of Plant Physiology, 96(1), 57-62 (2014)
Flow cytometric immunobead assay for the detection of BCR-ABL fusion proteins in leukemia patients
Weerkamp F. et al.
Leukemia, 23(6), 1106-1117 (2009)
Related Content
Protein purification techniques, reagents, and protocols for purifying recombinant proteins using methods including, ion-exchange, size-exclusion, and protein affinity chromatography.
Read More
Protein expression technologies for expressing recombinant proteins in E. coli, insect, yeast, and mammalian expression systems for fundamental research and the support of therapeutics and vaccine production.
Read More
Pull-down assays, reagents, and protocols for investigating in vitro protein-protein interactions using affinity or GST pull-down, tandem affinity purification (TAP), and co-immunoprecipitation methods.
Read More

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

Contact Technical Service

Social Media

LinkedIn icon
Twitter icon
Facebook Icon
Instagram Icon


Research. Development. Production.

We are a leading supplier to the global Life Science industry with solutions and services for research, biotechnology development and production, and pharmaceutical drug therapy development and production.

© 2021 Merck KGaA, Darmstadt, Germany and/or its affiliates. All Rights Reserved.

Reproduction of any materials from the site is strictly forbidden without permission.