Isolation of Centrosomal Fractions for Proteomic Studies in Lymphocytes

Centrosomes are essential microtubule-organizing centers that coordinate spatial cues for cellular polarization and organelle trafficking. In T lymphocytes, centrosomes translocate toward the immunological synapse to orchestrate polarized secretion and sustain signaling with the antigen-presenting cell. Understanding the dynamic composition of the centrosome during lymphocyte activation is crucial for uncovering the molecular mechanisms behind immune responses. Here, we describe a detailed protocol to isolate centrosome-enriched fractions from human T lymphoblasts under near-physiological conditions, optimized for downstream applications such as proteomic or structural analyses.