Knowledge Insight: Principles and Application Strategies of His-Tagged Protein Purification Technolog
Knowledge Insight: Principles and Application Strategies of His-Tagged Protein Purification Technology
Concept
The His-tag is a short polypeptide consisting of 6–10 consecutive histidine residues, genetically fused to the N- or C-terminus of a target recombinant protein. His-tag purification relies on immobilized metal affinity chromatography (IMAC), in which histidine imidazole groups coordinate with divalent metal ions (Ni²⁺, Co²⁺, Cu²⁺) to enable highly specific, efficient single-step protein purification.
Research Frontier
His-tag purification is the most widely used recombinant protein preparation method in modern biotechnology. Current advances focus on high-selectivity resins, automated process integration, high-throughput parallel purification, and green reusable materials, supporting structural biology, antibody drug development, vaccine research, and proteomic applications.
Research Significance
His-tag technology enables:
- Fast, mild, and cost-effective purification of recombinant proteins
- Compatibility with native or denaturing conditions
- Minimal impact on protein structure and bioactivity
- Seamless connection to functional analysis, crystallization, and drug discovery
- Standardized production for industrial and academic research
Core Mechanisms and Technical Methods
Key Advantages of His-Tag System
- Simple & rapid: One-step purification under native or denaturing conditions
- Highly versatile: Works in prokaryotic and eukaryotic expression systems
- Excellent compatibility: Suitable for cleavage, functional assays, and structural studies
Purification Principle
- Imidazole groups in His-tags form reversible coordination bonds with immobilized transition metals
- Binding occurs at pH 7.0–8.0; elution via competitive imidazole or low pH
- Metal selection: Ni²⁺ (high capacity), Co²⁺ (high selectivity), Cu²⁺ (strong affinity)
Common Purification Resins
- Pre-chelated resins: Ni-based (high capacity), Co-based (high selectivity)
- Self-chelating resins: IDA (tridentate), tetradentate (stable, harsh conditions)
Process Optimization
- Lysis: 20–50 mM imidazole to reduce non-specific binding
- Wash: 50–100 mM imidazole
- Elution: 150–500 mM imidazole
- Polishing: Desalting, tag cleavage, gel filtration
Typical Applications
- Vaccine antigen production (e.g., SARS-CoV-2 RBD)
- Antibody fragment (Fab, scFv) preparation
- Structural biology and crystallography
- Protein–protein interaction research
- High-throughput protein expression screening
Product Empowerment
ANT BIO PTE. LTD. provides the His-Tag Recombinant Rabbit Monoclonal Antibody (S-1398-151, Cat. No. S0B1838) under its Starter product line. This high-specificity, high-affinity reagent supports reliable detection and quality control of His-tagged proteins throughout expression, purification, and functional analysis.
Core Product Advantages
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Core Product Advantages |
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High Specificity & Superior Affinity: Specifically recognizes 6×His tags with minimal background; strong binding enables reliable detection of low-abundance proteins. |
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Excellent Stability & Batch Consistency: Strict QC ensures high stability and low inter-batch variation for reproducible long-term studies. |
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Broad Platform Compatibility: Validated for WB, IF, IHC, ELISA, and IP; ideal for protein expression verification and interaction analysis. |
Key Application Scenarios
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Key Application Scenarios |
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Recombinant Protein Detection: WB, IF, and IHC identification of His-tag fusion proteins. |
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Protein Purification QC: Monitoring expression levels and purification efficiency. |
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Protein Interaction Research: Capture or detection antibody in Co-IP assays. |
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High-Throughput Screening: ELISA and microarray-based quantification of recombinant proteins. |
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Subcellular Localization: Tracking His-tagged protein distribution in cells and tissues. |
Brand Mission
ANT BIO PTE. LTD. is a leading provider of life science reagents, offering antibodies, recombinant proteins, assay kits, and general laboratory reagents. We operate three specialized sub-brands: Absin (general reagents & kits), Starter (antibodies), and UA (recombinant proteins), delivering integrated solutions for protein science, recombinant expression, structural biology, and biopharmaceutical development. We are committed to supplying high-quality, consistent, and innovative tools to accelerate protein research and therapeutic innovation.
Related Product List
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Product Name |
Catalog No. |
Applications |
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His-Tag Recombinant Rabbit Monoclonal Antibody (S-1398-151) |
S0B1838 |
WB, IF, IHC, ELISA, IP |
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His-Tag Protein Purification Resin Kits |
Absin series |
IMAC purification of His-tag proteins |
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Recombinant His-Tag Control Protein |
UA-series |
Positive control and validation |
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Protein Lysis & Purification Buffers |
Absin series |
Sample preparation and stabilization |
Brand Promotion Copy
ANT BIO PTE. LTD. – Empowering Scientific Breakthroughs
At ANTBIO, we are committed to advancing life science research through high-quality, reliable reagents and comprehensive solutions. Our specialized sub-brands (Absin, Starter, UA) cover a full spectrum of research needs, from general reagents and kits to antibodies and recombinant proteins. With a focus on innovation, quality, and customer-centricity, we strive to be your trusted partner in unlocking scientific mysteries and driving medical progress. Explore our product portfolio today and elevate your research to new heights.
