The Protein Corner
This case study examines the Staley Lab’s success in using the CL7/Im7 system for their protein purification needs. Cody Hernandez, PhD candidate, shares his experience working with TriAltus’ methods to purify helicases, a mutant helicase, and several proteins.
Cas9 requires high purity in order to achieve high activity as a part of the CRISPR system for genetic modification. Due to a growing demand for Cas9 and an effort to improve its production, TriAltus conducted its own purification runs of Cas9 with success.
In the past several years, CRISPR has emerged as a powerful system for targeted gene modification. The system requires only two primary components: a guide RNA (gRNA or sgRNA) sequence and a Cas9 enzyme.
This blog post about pH in the context of protein purification is the third in a series about optimizing conditions for protein purification. Although pH is most commonly seen as an issue in ion-exchange chromatography, it also plays a critical role in affinity tag protein purification.
Using denaturing conditions is a way to coax insoluble proteins into solution by reducing hydrophobic effects and unfolding the aggregates. Denaturing agents are also useful for testing protein folding dynamics, protein elution from a column, and regenerating a resin column.
When conducting affinity purification of a protein, there are many variables to consider in order to optimize purity and yield. One such condition to control is high salt loading, which decreases impurities in the final product.
Protein fusion tags make affinity chromatography possible for many proteins, particularly those for which no affinity-quality antibody is available. In addition to improving conditions for purification, tags can facilitate detection and protein interaction studies.
Affinity purification is a powerful method for isolating proteins. In addition to providing affinity to separate proteins from solution, tags can have a profound impact on a protein’s folding or activity.
Protein purification can be a daunting topic. There are multitudes of methods available, each with their own set of conditions, requirements, and potential customizations.