Ex-Vivo Signal Transduction Studies in Chronic Lymphocytic Leukemia

Microenvironmental signaling is pivotal to chronic lymphocytic leukemia (CLL) pathology; therefore understanding how to investigate this pathway by both protein and chemical methods is crucial if we are to investigate and correlate biological changes with therapeutic responses in patients. Herein, we describe the use of western blotting also referred to as immunoblotting as a method that can semiquantitatively evaluate changes in protein expression following receptor engagement; this includes B cell receptor (BCR) signaling following stimulation with anti-IgM (Blunt et al. Clin Cancer Res 23(9):2313–2324, 2017). It is important to note that immunoblotting should always be combined with other quantitative methods such as flow cytometry to confirm activation of these signaling pathways (Aguilar-Hernandez et al. Blood 127(24):3015–3025, 2016).