MtoZ Biolabs' His Tagged Protein Purification Kit (Inclusion Body Protein) is designed to solubilize and purify His tagged recombinant proteins from bacterial inclusion bodies under denaturing conditions for downstream SDS-PAGE, Western blotting, activity assays, and LC-MS analysis.
Product Overview
Purifying His tagged proteins from inclusion bodies is challenging and often leads to low yield or poor purity. Key difficulties include:
1. Highly Aggregated Inclusion Bodies
Target proteins in inclusion bodies are tightly packed and misfolded, frequently requiring strong denaturants to fully solubilize the aggregates and expose the His tag for efficient binding to the metal affinity resin.
2. Co-Purification of Host Contaminants
Bacterial lysates from inclusion body preparations contain host proteins, membrane fragments, and endogenous histidine rich proteins that can bind nonspecifically to Ni or Co-based resins and reduce the purity of the His tagged product.
3. Viscous, Difficult to Handle Lysates
Cell disruption in high density cultures releases large amounts of genomic DNA and other macromolecules, creating viscous lysates that are hard to pipette, slow to clarify, and prone to trapping insoluble material.
4. Transition from Denaturing to Functional Buffers
Proteins purified from inclusion bodies must often be transferred from high denaturant conditions to refolding or assay buffers, and poorly controlled steps can cause aggregation and loss of usable protein.
Advantages
✔️Robust Solubilization of Inclusion Body Protein
MtoZ Biolabs' His Tagged Protein Purification Kit (Inclusion Body Protein) provides denaturing lysis and binding buffers formulated to efficiently disperse inclusion bodies and keep His tagged proteins soluble, improving capture efficiency on the affinity resin.
✔️Improved Selectivity for His Tagged Targets
Optimized wash and elution conditions help reduce retention of host contaminants and histidine rich background proteins, delivering higher purity His tagged protein than generic Ni-NTA workflows.
✔️Reduced Lysate Viscosity for Easier Processing
The extraction system is designed to limit DNA-driven viscosity and facilitate clarification, yielding lysates that are easier to centrifuge, filter, and load onto columns or magnetic formats.
✔️Refolding Ready Eluates
MtoZ Biolabs' His Tagged Protein Purification Kit (Inclusion Body Protein) produces denatured but well defined His tagged protein fractions that are compatible with downstream buffer exchange and refolding screens, supporting reliable structural and functional 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.
