MtoZ Biolabs Kinase Enrichment Kit with ADP Probe - A Proteomics Platform Based on ActivX ADP Probes That Covalently Captures and Analyzes the Functional Kinome, Enabling Drug Discovery from Activity Profiling to Precise Target Identification
Product Overview
1. Inability to Directly Assess Kinase Activation States
Techniques such as Western blotting primarily report the presence or expression levels of kinase proteins, but do not provide information on whether the kinases adopt catalytically competent, active conformations.
2. Labor-Intensive and Low-Throughput Inhibitor Screening and Validation
Assessment of binding affinity and target occupancy for small-molecule inhibitors typically relies on individual biochemical or cellular assays tailored to specific kinases, resulting in limited throughput and reduced capability to identify previously unrecognized off-target proteins.
3. Challenges in Comprehensive Off-Target Profiling
Hypothesis-driven validation strategies (e.g., assays focused on known targets) may overlook critical off-target interactions, leading to an incomplete characterization of drug mechanisms of action and potential toxicological liabilities.
4. Limited Coverage of Inactive Kinase States
A substantial fraction of kinases retain the ability to bind nucleotides or inhibitors even in inactive conformations; however, traditional activity-based enrichment strategies may not efficiently capture this class of kinases, thereby missing important regulatory and mechanistic information.
Advantages
✅ Active-Site-Directed Covalent Labeling
The core component of the MtoZ Biolabs Kinase Enrichment Kit with ADP Probe, the ActivX desulfurized biotin ADP probe, is a nucleotide-derived molecule that selectively and covalently modifies the conserved lysine residue within the kinase active site, thereby directly labeling catalytically competent nucleotide-binding pockets.
✅ Simultaneous Profiling of Active and Inactive Enzymes
The unique probe design allows binding and labeling of both active and inactive enzymes present in the sample, enabling a more comprehensive characterization of kinase functional states and offering a unique perspective for investigating the regulation of enzymatic activity.
✅ Flexible Competitive Binding and Elution Strategy
Co-incubation with competitive small-molecule inhibitors enables precise quantification of inhibitor binding affinity. Owing to the relatively weak interaction between desulfurized biotin and streptavidin, elution can be performed under mild and reversible conditions, thereby maximally preserving protein and peptide integrity and ensuring compatibility with downstream analyses.
✅ Robust Compatibility with Dual Downstream Analytical Workflows
Enriched desulfurized biotin-labeled proteins can be subjected to SDS-PAGE analysis, followed by validation of known targets using specific antibodies, making this approach suitable for Western blotting (WB). In parallel, enriched and proteolytically digested desulfurized biotin-labeled peptides can be analyzed by LC-MS/MS, enabling global and unbiased identification of dozens to hundreds of kinases and other ATPases. This strategy represents a gold standard for the discovery of novel targets and off-target effects in mass spectrometry-based studies.
Technical Support
1. Protocol Downloads
MtoZ Biolabs provides comprehensive experimental protocols and product manuals to help ensure smooth and reproducible workflows. For access to specific documents or detailed instructions, please contact our support team.
2. Customized Service Support
Our experienced scientists offer tailored technical assistance and customized solutions based on your specific research goals, helping you achieve optimal experimental outcomes.
3. FAQ and Expert Assistance
We provide responsive and professional technical support to address your questions efficiently. Our experts are ready to help you identify and resolve experimental challenges quickly and accurately.
