Multiplexed isobaric tagging protocols for quantitative mass spectrometry approaches to auditory research

Area of Science:

• Proteomics
• Molecular Biology
• Biochemistry

Background:

• Biologists utilize numerous techniques for molecular quantification.
• Implementing these techniques can be challenging, requiring careful protocol selection and validation.
• Choosing the right protocol involves assessing desired results, data interpretability, and sample compatibility.

Purpose of the Study:

• To detail a robust protein labeling procedure for differential quantification of protein abundance.
• To provide a method applicable to complex cell lysates and adaptable for other tissues.
• To guide researchers in selecting appropriate protein labeling strategies for mass spectrometry.

Main Methods:

• A detailed protocol for labeling proteins in complex cell lysates.
• Adaptations for labeling lysates from various tissues.
• Mass spectrometric analysis for peptide quantification and protein identification.

Main Results:

• Successful implementation of a protein labeling procedure.
• Generation of interpretable and unbiased data for differential protein quantification.
• Enabling accurate protein identification and abundance measurement.

Conclusions:

• The presented protein labeling method is robust and effective for differential quantification.
• The protocol is adaptable for various biological samples, including cell cultures and tissues.
• This work facilitates accurate proteomic analysis through mass spectrometry.