Protein Degradation or Enzyme Digestion Analysis Service

Protein degradation is crucial for regulating protein function and levels within cells. Specific proteases cleave proteins into smaller fragments, impacting their structure and function, and generating biologically active cleavage products. Understanding degradation pathways and cleavage sites is essential for studying protein function, signal transduction, and metabolic regulation. Enzymatic digestion analysis uses specific proteases (e.g., trypsin, staphylococcus protease) to degrade proteins into peptides suitable for mass spectrometry analysis. These peptides' mass spectrometry data facilitate protein identification, sequence determination, and post-translational modification studies.

 

In modern life sciences, protein degradation and enzymatic digestion analysis are indispensable. They elucidate protein structure and function and offer insights into dynamic biological processes. MtoZ Biolabs leverages advanced mass spectrometry and extensive proteomics expertise to deliver efficient, precise protein degradation and enzymatic digestion analysis. Free project evaluation, welcome to learn more details! Our technical specialists are available to provide a free business assessment.

 

Technical Principles

1. Sample Preparation

(1) Sample extraction: Extracting proteins from biological samples such as cells, tissues, or serum and using a protein extraction buffer to ensure protein stability during extraction.

(2) Protein quantification: Quantifying protein concentration using methods such as the BCA assay, Bradford assay, or UV absorbance to ensure that the protein content in the sample is suitable for subsequent analysis.

 

2. Protein Degradation

(1) Reduction and alkylation: Reducing disulfide bonds in the protein sample (typically using dithiothreitol (DTT) or tris(2-carboxyethyl)phosphine (TCEP)) and performing alkylation to prevent the reformation of disulfide bonds (typically using iodoacetamide or iodoacetic acid).

(2) Enzymatic digestion: Selecting an appropriate protease (such as trypsin, lysozyme, PNGase F, etc.) for digestion and mixing the protein sample with the enzyme according to the enzyme's instructions, followed by incubation at the appropriate temperature and time (e.g., trypsin is usually incubated at 37 ℃ for 16 hours).

 

3. Separation and Purification of Digestion Products

Purifying digestion products using solid-phase extraction (SPE) columns, reverse-phase high-performance liquid chromatography (RP-HPLC), or other methods to remove impurities and non-target peptides and collecting target peptides for further analysis.

 

4. Analysis and Identification

(1) Mass spectrometry analysis: Performing mass spectrometry analysis (such as liquid chromatography-mass spectrometry (LC-MS/MS)) on the purified peptide samples. 

(2) Data analysis: Analyzing mass spectrometry data using specialized software (such as Mascot, Sequest, MaxQuant, etc.) to match known protein sequences in databases to identifyi peptides and proteins and performing quantitative analysis (such as label-based quantification, label-free quantification) to compare protein expression levels between different samples.

 

5. Report Generation

Interpreting the results of protein degradation or enzymatic digestion analysis based on mass spectrometry data and software analysis, determining protein sequences, modifications, and functions and generating a detailed analysis report.

 

6. Experimental Validation

Using other methods (such as Western blot, ELISA, etc.) to validate mass spectrometry analysis results, ensuring data accuracy and reliability.

 

Service Advantages

1. Expert Team

Our scientists and technicians have extensive proteomics knowledge and mass spectrometry analysis experience.

 

2. Rapid Turnaround

We ensure fast delivery, typically completing analyses within a few weeks, without compromising quality.

 

3. Customized Service

We offer tailored analysis services and detailed reports to meet customer needs, solving complex research challenges.

 

Deliverables

1. Experimental Procedures

2. Relevant Mass Spectrometry Parameters

3. Detailed Information on Protein Degradation or Enzyme Digestion Analysis

4. Mass Spectrometry Images

5. Raw Data