3.3 Western blotting
Western blotting is a widely used technique to detect target protein presence, size, abundance, and modifications in research. It’s valuable for diagnostics, therapeutics, and academic studies. This method transfers protein patterns from a gel to a membrane and has evolved since its introduction in 1979 [32]. Western blotting is also essential for identifying microorganism-related antigens and infectious diseases. For example, it assesses HSV-2 seroprevalence by measuring specific immunoglobulin G levels [33] and identifies Leis mania donovani through Hsp83 and Hsp70 antigen detection [34]. This technique has diverse applications, including protein quantity, kinase function, cellular localization, protein-protein interactions, and post-translational modifications. Its use in skeletal muscle and exercise physiology research is growing, but maintaining quality control is crucial to avoid misleading data [35].
Cell uniqueness is a fundamental aspect of biology, impacting development, stem cell biology, and cancer. Conventional bulk measurements, however, mask the intricate biology within individual cells. Single-cell measurement techniques are essential to unveil cell-to-cell variation, and microfluidic instruments are indispensable for precise biochemical experiments at this level. Single-cell western blotting (scWB), akin to traditional western blotting, proves especially advantageous when dealing with protein targets lacking specific antibodies or when background signal from intact cells poses challenges. This method is invaluable for direct protein quantification in single cells, with applications spanning basic bioscience research to practical biomedical purposes [36].