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An increasingly useful technique in the study of biological systems is proteomics. It is possible to identify the cellular and molecular mechanisms involved in a biological process by comparing two different physiological states. Based on the mRNA content of proteins, proteomics can confirm their presence and provide a precise estimate of how abundant they are in a cell. Proteomic techniques can be employed on a global or targeted basis. Targeted proteomics methodologies restrict the number of characteristics to be monitored before optimizing the methods to get the maximum sensitivity and throughput for a large volume of data. Global proteomics approaches have the advantage of requiring just a discernible variation in one or more protein species between samples, negating the need for a hypothesis. The goal of global proteomics techniques is to separate, quantify, and identify every protein present in a sample. This study focuses solely on the various methodologies for protein and peptide separation and quantification to conduct a comparative and quantitative global proteomics investigation. Protein quantification will be covered both in-gel and off-gel, as well as the accompanying mass spectrometry technique. The review focuses on the most common strategies, bearing in mind that each approach is modular and frequently recovers the other.