Lactylation analysis is particularly relevant for systems involving lactate accumulation, enhanced glycolysis, hypoxic conditions, tumor microenvironment studies, immune cell activation, or metabolic reprogramming. If your research touches on these processes, it is highly likely that lysine lactylation plays a regulatory role and can be effectively profiled using lactylation proteomics.

Not necessarily. Basal lactylation levels can be detected in many cell types and tissues without additional treatment. However, conditions such as lactate supplementation, hypoxia, glycolysis activation, oxidative phosphorylation inhibition, or inflammatory stimulation often enhance lactylation signals and help reveal dynamic regulatory changes.

Yes, lactylation is a low-abundance and sample-quality–dependent post-translational modification. To preserve lactylation sites, it is critical to process samples rapidly at low temperature, avoid repeated freeze–thaw cycles, use protease and deacylase inhibitors, and minimize processing time. Proper sample handling significantly improves detection sensitivity.

Lactylation generally has lower abundance, higher background complexity, and stronger dependence on metabolic states, making detection more demanding. Therefore, high-specificity antibody enrichment combined with highly sensitive LC-MS/MS acquisition is essential to achieve reliable site-level identification.

Yes. Lactylation is often more pronounced in highly glycolytic or metabolically active tissues/cells, such as: Tumor tissues and cancer cell lines; Activated immune cells (e.g., macrophages, dendritic cells); Tissues under hypoxia or inflammation; Energy-demanding organs like the brain and muscle. These sample types typically yield stronger lactylation signals.

Yes. While each PTM requires separate specific enrichment, lactylation proteomics can be combined with acetylation, succinylation, malonylation, and other metabolic acylations to explore cross-talk between metabolic pathways and epigenetic regulation. Multi-PTM analysis is particularly informative for studying metabolic–epigenetic interactions.

Generally, lactylation increases with elevated lactate, but the relationship is not strictly linear. Lactylation depends on multiple factors, including lactyl-CoA availability, lactate metabolism, enzyme activities, and substrate accessibility. As such, lactylation reflects a composite regulatory response rather than a direct one-to-one readout of lactate concentration.