Histone Post-Translational Modification Analysis

BTP-Analysis of Histone Post-Translational Modifications

Histones are a class of highly conserved proteins, with only a few amino acid differences between different variant sequences. The N-terminus of histones can undergo various modifications such as methylation, acetylation, phosphorylation, ubiquitination, and ADP-ribosylation. These covalent modifications are closely related to gene transcription, DNA damage repair, DNA replication, and chromosome condensation. As an important epigenetic marker, histone modifications also have certain connections with other epigenetic markers, forming a complex network. The histone code greatly enriches the informational content of the traditional epigenetic code.

BTP offers a perfect solution for qualitative and quantitative analysis of histone modifications using Thermo Fisher's Q ExactiveHF mass spectrometry platform, Orbitrap Fusion mass spectrometry platform, and Orbitrap Fusion Lumos mass spectrometry platform combined with Nano-LC. Simply tell us your experimental objectives and send us your samples, and we will handle all subsequent aspects of the project, providing identification and quantitative analysis of histone modification types and sites.

Compared to traditional quantitative proteomic analysis, a key step in quantitative analysis of histone modifications is how to isolate and purify pure histone components. Since histones are located in the cell nucleus, based on an article published in Nature Protocol titled: 'Extraction, purification and analysis of histone', BTP has established an experimental platform for histone isolation and purification by combining its own experimental experience. This involves cell homogenization and low-speed centrifugation to obtain the nucleus, followed by further nucleus disruption, separation of histones and DNA, precipitation of impurities with high-salt buffer, and HPLC purification to obtain various histone components for subsequent mass spectrometry analysis. Additionally, to identify and quantify more histone post-translational modification information, we use 2-3 different enzymes for protein digestion during the enzymatic digestion process, improving peptide coverage in mass spectrometry analysis and preventing the loss of histone post-translational modification information due to inappropriate peptide length and low ionization efficiency.

Research Route for Identification of Histone Post-Translational Modifications

BTP-Sample Requirements for Histone Post-Translational Modifications Analysis

1. If you provide tissue samples, please send them on dry ice. Plant tissue samples should be more than 400mg, blood samples at least 2ml (use EDTA for plasma anticoagulation), serum 2ml, urine 10ml, animal tissue samples at least 2g, cell samples should be 1X10^8 cells, yeast and microbial samples should have a dry weight of 400mg.
2. If you provide protein samples, please ensure the total protein amount is between 2-5mg. Ordinary tissue and cell lysis buffers can be used for protein extraction.
3. Sample transportation: Please use sufficient dry ice for transportation and choose a fast delivery method as much as possible to minimize the possibility of sample degradation during transportation.
4. Before the formal experiment, we will test the samples you provide, and the formal experiment will begin once the samples pass the test.

BTP-Research Case Studies for Histone Post-Translational Modifications Analysis

Identification of Ubiquitination Sites on Proteins

Bilingual Project Report in Chinese and English

In the technical report, BTP will provide you with a detailed bilingual report in Chinese and English, which includes:

1. Experimental Procedures (Chinese and English)
2. Relevant Mass Spectrometry Parameters (Chinese and English)
3. Detailed Information on Identified Histone Post-Translational Modifications
4. Mass Spectrometry Images
5. Raw Data

BTP-One-Stop Service for Histone Post-Translational Modifications Analysis

You only need to place an order and send your samplesBTP One-Stop ServiceCompletion: Sample Processing - Instrument Analysis - Data Analysis - Project Report

Related Services

Proteomics Analysis of Post-Translational Modifications
Phosphorylation Quantitative Proteomics Research
Acetylation Quantitative Proteomics Research
Ubiquitination Quantitative Proteomics Research
Glycosylation Quantitative Proteomics Research
Methylation Quantitative Proteomics Research
Peptide Coverage/Peptide Spectrum Analysis