Sigma introduces one-step affinity purification column

By Wai Lang Chu

- Last updated on GMT

Related tags Protein Molecular biology Dna

Sigma Aldrich has launched a new integrated lysis and affinity
purification (iLAP) column, which greatly simplifies the process of
cell lysis and histidine-tagged protein purification into one step,
allowing fast, efficient and non-denaturing protein extraction
directly from microbial cultures.

The technology allows for simultaneous cell lysis and protein purification, which will reduce purification time as well as sample handling, minimising the potential for protein loss or damage.

Analysts have been commenting in recent years that the use of recombinant tagged proteins will increase further since functional biology and structural genomics now are "booming."

The Histidine-tag is small and does not, in most cases, interfere with the function/activity/structure of the protein, but to cleave off the tag is becoming more and more common.

There has now become a need to increase the throughput in protein purification and using tags achieves this by adding simplicity to the purification protocol.

Sigma's HIS-Select iLAP column 5 ml Column consists of a novel single-use, disposable column designed for the one-step purification of histidine-tagged proteins directly from a 5 ml bacterial culture.

This product maximises recombinant protein extraction while eliminating sample loss. The column contains highly specific HIS-Select nickel chelate matrix and highly efficient CelLytic Express, both present in a convenient tablet format.

The column is designed for the simultaneous, direct lysis of a 5 ml bacterial culture and affinity capture of the target histidine-tagged protein without any additional manipulation required (such as centrifugation).

As the culture is lysed in the column, the target protein is captured on the same highly selective HIS-Select Affinity Gel. The relatively high capacity feature of these columns enables capture of at least 1-2 mg of histidine-tagged protein per column, making them ideal for confirming expression of a histidine-tagged target protein, for performing protein:protein interaction assays, or for rapidly screening colonies or multiple constructs (cultures).

The resulting purified protein is compatible with protein assays, SDS-PAGE, Western blotting and MALDI.

"This new 5 ml column format allows for the rapid screening of recombinant clones for protein expression and subsequent protein characterisation,"​ Michael Hadjisavas, Global Strategic marketing manager of Proteomics, at Sigma-Aldrich Corporation.

"The HIS-Select iLAP technology has simplified purification of histidine-tagged recombinant proteins directly from bacterial cultures,"​ he added.

Sigma's introduction of this new technology brings it up against the likes of Amersham Bioisciences, Roche, and Clontech, who all make available products for the purification of histidine-tagged recombinant proteins.

For more information about the HIS-Select iLAP column 5 ml Column visit Sigma Aldrich's website.

Related topics Preclinical Research

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