2-Hydroxyisobutyrylation Analysis Service

2-Hydroxyisobutyrylation (Khib) is an emerging post-translational modification characterized by the covalent attachment of a 2-hydroxyisobutyryl group to lysine residues. This acylation event alters the electrostatic and structural properties of proteins, thereby affecting chromatin architecture, transcriptional regulation, and metabolic control. Because of its roles in diverse cellular pathways and disease mechanisms, Khib has become a rapidly expanding research focus in both proteomics and epigenetics.

MtoZ Biolabs provides a dedicated 2-Hydroxyisobutyrylation Analysis Service that includes two complementary analytical options: target protein 2-hydroxyisobutyrylation analysis and 2-hydroxyisobutyrylation proteomics. By combining selective enrichment, high-resolution mass spectrometry, and advanced bioinformatics pipelines, our platform supports both focused and global investigations into Khib-mediated regulation.

1. Target Protein 2-Hydroxyisobutyrylation Analysis

This service is tailored for researchers investigating Khib on specific proteins of interest. Through optimized antibody-based or affinity capture strategies combined with LC–MS/MS, MtoZ Biolabs accurately identifies 2-hydroxyisobutyrylation sites and quantifies modification levels under different biological conditions. This targeted approach enables detailed functional assessment of individual proteins, such as histones, metabolic enzymes, or signaling regulators, revealing how Khib modulates their structural conformation, enzymatic activity, and interaction networks.

2. 2-Hydroxyisobutyrylation Proteomics

For discovery-driven studies, MtoZ Biolabs offers 2-hydroxyisobutyrylation proteomics, which provides large-scale profiling of Khib modifications across the proteome. Using high-throughput sample preparation, specific peptide enrichment, and high-resolution LC–MS/MS acquisition with quantitative options (label-free, TMT, or SILAC), we comprehensively map modification sites and compare Khib patterns across tissues, developmental stages, or treatment groups. Downstream bioinformatics analysis further integrates pathway enrichment, network modeling, and cross-talk evaluation with other lysine acylations to deliver system-level insight into Khib regulation.

What is 2-Hydroxyisobutyrylation?

Protein post-translational modifications (PTMs) expand the functional diversity of the proteome by regulating activity, localization, and interactions of proteins. Among these, lysine acylations such as acetylation, crotonylation, and succinylation have been extensively studied. Lysine 2-Hydroxyisobutyrylation (Khib) represents a relatively novel addition to this group, characterized by the attachment of a 2-hydroxyisobutyryl moiety to lysine residues.

This modification introduces bulkier and more hydrophilic side chains compared to acetylation, which can lead to distinct biochemical effects on proteins. It has been reported to regulate chromatin structure and gene transcription, particularly through histone modification. For example, histone 2-hydroxyisobutyrylation is linked to active gene expression, suggesting its role as an epigenetic marker. Beyond histones, non-histone proteins involved in metabolism and signaling have also been identified as substrates, indicating broad regulatory functions. Dysregulated 2-hydroxyisobutyrylation has been associated with cancer, metabolic disorders, and stress responses, highlighting its clinical relevance.

Analysis Workflow

Bo, F. et al. Sci Rep. 2021.

Figure 1. Systematic Analysis of Khib in Wheat Roots

1. Protein Extraction

Proteins are extracted from tissues, cultured cells, or biofluids under optimized conditions to maintain modification stability and sample quality.

2. Enzymatic Digestion

Extracted proteins are digested into peptides using high-grade proteases to enable downstream site-specific analysis.

3. HPLC Fractionation

Peptides are fractionated by high-performance liquid chromatography (HPLC) to reduce complexity and improve coverage of modified peptides.

4. Enrichment of Modified Peptides

Specialized enrichment strategies, such as antibody-based capture, are applied to selectively isolate Khib-modified peptides from the peptide pool.

5. LC-MS/MS Analysis

Enriched peptides are analyzed on high-resolution LC-MS/MS platforms, such as Orbitrap or Q Exactive HF, for confident identification of modification sites.

6. Data Analysis

Mass spectrometry data are processed and interpreted through robust pipelines, including peptide-spectrum matching, site localization, functional annotation, pathway mapping, and network analysis.

7. Report Delivery

Clients receive a publication-ready report containing experimental details, identified sites, functional analyses, visualizations, and both raw and processed datasets.

Sample Submission Suggestions

1. Sample Types

We accept various biological sample types, including but not limited to:

• Cultured cells
• Microorganisms
• Animal or human tissue
• Plant tissue
• Biological fluids such as plasma or serum
• Protein Extracts

2. Storage and Transport

• Samples should be snap-frozen in liquid nitrogen and stored at –80°C until shipment.
• Ship samples on dry ice.
• Avoid repeated freeze–thaw cycles.

*Note: If you have special sample types or require additional guidance, please contact us for personalized support before sample preparation.

Service Advantages

✔️Advanced Analytical Platforms

Utilization of cutting-edge LC-MS/MS systems with optimized enrichment technologies.

✔️Experienced Team

Extensive expertise in PTM proteomics, bioinformatics, and functional interpretation.

✔️Comprehensive Approach

From global profiling to targeted validation, we provide end-to-end solutions.

✔️Customizable Solutions

Flexible service options tailored to meet specific client requirements.

✔️High-Quality Data

Stringent quality control ensures reproducibility and publication-ready results.

✔️Rapid Turnaround

Efficient workflows and project management enable timely delivery.

FAQ

Q1: Can Khib analysis be integrated with other PTM studies such as acetylation or succinylation?

A1: Yes. For clients interested in multi-PTM profiling, we can design workflows that combine Khib analysis with other lysine acylations to provide a broader regulatory landscape.

Q2: How do you ensure specificity during Khib peptide enrichment?

A2: We employ antibody-based enrichment validated for Khib specificity, along with optimized washing and control conditions, to minimize non-specific binding and ensure reliable results.

Deliverables

1. Comprehensive Experimental Details

2. Materials, Instruments, and Methods

3. The Detailed Information of 2-Hydroxyisobutyrylation Analysis

4. Mass Spectrometry Image

5. Bioinformatics Analysis

6. Raw Data

Our comprehensive 2-Hydroxyisobutyrylation Analysis Service supports research in epigenetics, disease mechanisms, and therapeutic development, delivering reliable data and meaningful interpretation. Contact us today to discuss your project and request a customized proposal!