Histone Crotonylation Analysis
- Animal tissue ≥ 2 g
- Plant tissue ≥ 0.4 g
- Serum / Plasma ≥ 2 mL (plasma should be collected using EDTA as an anticoagulant)
- Urine ≥ 10 mL
- Cell ≥ 1 x 10⁸ cells
- Yeast / Microbial samples ≥ 0.4 g
- Protein samples: 2-5 mg (Standard tissue or cell lysis buffers can be used during protein extraction)
Histone crotonylation is a critical post-translational modification that involves the addition of a crotonyl group to lysine residues on histone proteins. This modification has significant implications for gene expression, chromatin dynamics, and cellular metabolism. Crotonylation is primarily catalyzed by histone crotonyltransferases, which utilize crotonyl-CoA as a substrate. Crotonyl-CoA is produced from crotonate, a short-chain fatty acid, through enzymatic pathways involving acyl-CoA synthetase. The balance between crotonyl-CoA and other acyl-CoA species, such as acetyl-CoA, is crucial for determining the degree of histone crotonylation, linking metabolic states to epigenetic regulation.
Aberrations in histone crotonylation have been implicated in various disease processes. For instance, in neuropsychiatric disorders, histone crotonylation influences synaptic plasticity by modulating the expression of neuropeptides such as VGF, thereby affecting mood and stress responses. In cardiovascular diseases, the hydrolysis of crotonyl-CoA by the enzyme ECHS1 is essential for maintaining normal crotonylation levels, which in turn regulates hypertrophic gene expression and cardiac remodeling. Furthermore, histone crotonylation has been identified as a critical factor in the latency and reactivation of HIV, with the enzyme ACSS2 playing a pivotal role in this process. Inhibiting ACSS2 can diminish histone crotonylation, subsequently reducing HIV replication. This highlights the potential of targeting histone modifications in therapeutic strategies for viral infections.
Figure 1. The Functions of Histone Crotonylation in Diseases
Services at MtoZ Biolabs
MtoZ Biolabs utilizes Thermo Fisher's Q Exactive HF, Orbitrap Fusion, and Orbitrap Fusion Lumos mass spectrometry platforms, in combination with Nano-LC, to provide a comprehensive solution for histone crotonylation analysis. Simply provide your experimental objectives and send us your samples, and we will offer you a one-stop service from sample preparation and MS analysis to data analysis, delivering detailed identification and quantitative analysis of histone crotonylation modifications and their modification sites.
Compared to traditional proteomics analysis, the key to histone modification analysis lies in the isolation and purification of histone components, as well as mitigating the impact of N-terminal lysine on the analysis of histone post-translational modifications. Since histones are located in the cell nucleus, MtoZ Biolabs has developed a specialized platform for histone extraction and purification, drawing on existing literature and experimental experience. Cells are homogenized and subjected to low-speed centrifugation to obtain the nuclei. The nuclei are then further disrupted to release histones and DNA. High-salt buffer is used to remove impurities, and High-Performance Liquid Chromatography (HPLC) is employed to separate different histone components, achieving optimal purity for subsequent mass spectrometry analyses. Additionally, to eliminate the impact of N-terminal lysine, we utilize 2-3 different enzymes to enhance peptide coverage in mass spectrometry analysis during the digestion process. This multi-enzymatic digestion increases the coverage of histone peptides, preventing the loss of post-translational modification information due to unsuitable peptide lengths or low ionization efficiency.
Analysis Workflow
Service Advantages
1. Advance Analysis Platform
Mtoz Biolabs established an advanced histone crotonylation analysis platform, guaranteeing reliable, fast, and highly accurate analysis service.
2. One-Time-Charge
Our pricing is transparent, no hidden fees or additional costs.
3. High-Data-Quality
Deep data coverage with strict data quality control. AI-powered bioinformatics platform integrate all histone crotonylation analysis data providing clients with acomprehensive data report.
4. Optimized Extraction
An optimized histone extraction and purification protocols are employed to minimize contamination and ensure high purity.
Sample Submission Requirements
1. Sample Volume
2. Sample Preservation
Samples should be shipped with dry ice. Avoid repeated cycles of freezing and thawing
Note: We will conduct testing on the provided samples. The formal experiment will commence only after the samples pass the quality check.
Deliverables
1. Comprehensive Experimental Details (Materials, Instruments, and Methods, etc.)
2. Relevant Liquid Chromatography and Mass Spectrometry Parameters
3. Detailed Information on Histone Crotonylation Analysis
4. Raw Data
5. Mass Spectrometry Image
6. Custom Analysis Report
MtoZ Biolabs, an integrated Chromatography and Mass Spectrometry (MS) Services Provider, provides advanced proteomics, metabolomics, and biopharmaceutical analysis services to researchers in biochemistry, biotechnology, and biopharmaceutical fields. Our ultimate aim is to provide more rapid, high-throughput, and cost-effective analysis, with exceptional data quality and minimal sample consumption. If you are interested in our histone crotonylation analysis services, please feel free to contact us. We are dedicated to providing you with the best service.
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