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].