Protein Enrichment

Protein enrichment is the process of increasing the concentration of a specific protein or group of proteins from a complex mixture, such as cell lysates or tissue extracts. This is often a preliminary step before protein characterization techniques, including mass spectrometry or western blotting. Numerous techniques are available for the isolation of cellular proteins based on their post-translational modification (PTM) or active binding sites. Protein enrichment, whether for glycoprotein purification, ubiquitin purification, kinase purification, or N-acetyl glucosamine enrichment is important for isolating low-abundance proteins or isolating protein classes. Enrichment of specific proteins or protein complexes can most easily be accomplished using target-specific immobilized antibody or PTM specific affinity binding. Protein enrichment techniques, such as immunoprecipitation, affinity purification, and chromatography, isolate and concentrate specific proteins from complex mixtures. These methods improve the accuracy and sensitivity of downstream analyses by enhancing the detection of low-abundance proteins. Protein enrichment is also important for analyzing complex samples as it enhances the detection of target proteins by concentrating them and reducing background noise. This step helps ensure accurate and reliable results in proteomics studies and diagnostic applications.

The novel Thermo Scientific ActivX™ probes and enrichment kits enable the specific targeting of several enzyme classes, including kinases, GTPases, and serine hydrolases.

• Comprehensive—reagents and kits for multiple protein enrichment strategies
• Optimized—all reagents and kits are designed to maximize protein yield and reduce nonspecific binding
• Convenient—easy-to-use spin formats enable more rapid enrichment of proteins
• Flexible—kits can be used with either cultured mammalian cells or tissues

• Phosphoprotein purification enables the selective enrichment of phosphorylated proteins or peptides to support signaling pathway analysis and phosphorylation-based regulation studies
• Glycoprotein purification isolates proteins with specific carbohydrate modifications, supporting characterization of protein structure, function, and cell–cell interactions
• Ubiquitin purification enriches ubiquitinated proteins or peptides to enable analysis of protein degradation, turnover, and ubiquitin-mediated signaling pathways
• Kinase purification supports the enrichment of active or total kinases for functional studies, inhibitor screening, and investigation of phosphorylation-driven signaling networks
• N-acetyl glucosamine enrichment selectively captures O-GlcNAc–modified proteins or peptides to support studies of post-translational regulation and cellular stress responses

For downstream mass spectrometry analysis, peptide enrichment and/or fractionation is often necessary to detect peptides from low-abundance proteins and/or for reducing sample complexity for proteomic analysis.

With the Pierce Phosphoprotein Enrichment Kit, achieve higher and faster phosphoprotein yields in less time.

*From 2 mg total protein.
**Based on maximum 1 mg load per manufacturer’s protocol.