Protein Digestion Service

Resolution and Specificity: It allows for precise analysis of proteins that have been separated into distinct bands or spots on a gel. This high-resolution separation helps in identifying individual proteins and characterizing their properties.

Reduced Complexity: By digesting proteins directly within the gel, you avoid issues related to sample handling and potential losses during transfer and processing.

Reproducibility: The process is well-established and reproducible, making it a reliable choice for detailed proteomics studies.

Speed and Efficiency: It is generally faster than in-gel digestion as it bypasses the gel electrophoresis step, making it suitable for larger sample numbers or when time is a constraint.

Flexibility: This method is versatile and can be used with a wide range of sample types, including those not separated by electrophoresis.

Ease of Implementation: The method involves fewer steps compared to in-gel digestion, which can simplify the workflow.

Each method has its limitations:

• In-Gel Digestion: While effective, this method is labor-intensive and time-consuming. It requires careful handling of gel slices and may be less practical for processing a large number of samples simultaneously.
• In-Solution Digestion: This approach may result in sample losses due to incomplete resolubilization of proteins, especially when dealing with proteins that aggregate or are poorly soluble in the urea solution. There can also be issues with contaminants if the initial precipitation step is not thorough.

Choosing the appropriate method depends on your experimental requirements:

In-Gel Digestion: Opt for this method if you have proteins that have been separated by gel electrophoresis and you need high-resolution analysis of specific bands or spots.

In-Solution Digestion: This method is preferable if you are dealing with complex mixtures that have not been previously separated or if you need a quicker, more streamlined digestion process.

Yes, both methods are designed to handle complex samples. In-gel digestion works well for samples that have been separated and need detailed analysis, while in-solution digestion can be applied to complex mixtures where separation is not a prerequisite. However, the effectiveness of digestion and the quality of data obtained can vary based on the sample type and the chosen method.

We implement stringent quality control measures throughout the digestion process:

• Standard Operating Procedures: We follow well-established protocols to ensure consistency and accuracy.
• Experienced Personnel: Our skilled team monitors each step, from sample preparation to digestion and analysis, to maintain high standards.
• Quality Checks: We perform regular checks and validations to ensure that the digestion process yields reliable and reproducible results.

For optimal results, please provide:

• Sample Information: Details about your protein samples, including concentration, buffer conditions, and any special handling requirements.
• Sample Format: Samples should be in a format compatible with the chosen digestion method (e.g., gel slices for in-gel digestion or solution for in-solution digestion).
• Specific Instructions: Any particular requirements or preferences for your study, such as desired protease or digestion conditions.

In-Gel Digestion: This method is suitable for proteins that have been separated by electrophoresis and are well-resolved into bands or spots. It is ideal for analyzing individual proteins in a complex mixture.

In-Solution Digestion: This method is more suitable for crude or complex protein mixtures that have not been separated. It works well for a wide range of proteins, including those that may aggregate or are otherwise challenging to handle.

For samples with high levels of contaminants, we perform additional purification steps before digestion. In the in-solution digestion method, we use chloroform/methanol precipitation to remove contaminants and concentrate the proteins. For in-gel digestion, we ensure that gel slices are thoroughly washed to remove residual contaminants. If necessary, we can employ additional clean-up procedures to ensure high-quality results.

The yield of peptides can vary depending on the protein's size, solubility, and the digestion method used. Generally, you can expect a high yield of peptides if the digestion process is optimized and if the proteins are well-solubilized. In-solution digestion typically results in a higher peptide yield compared to in-gel digestion, due to fewer losses during sample handling.

Yes, we can handle proteins with post-translational modifications (PTMs). Both in-gel and in-solution digestion methods are compatible with PTMs, although the choice of digestion conditions might be adjusted to optimize the recovery and analysis of modified peptides.