MLCK Inhibitor peptide 18 is a cell permeable inhibitor of myosin light chain kinase (MLCK; IC50 = 50 nM) with 4,000-fold selectivity for MLCK over CaM kinase II and PKA. It prevents defects in transepithelial barrier resistance induced by bacteria or TNF-α plus IFN-γ. MLCK Inhibitor peptide 18 prevents sperm chromatin-induced cortical reorganization in mouse oocytes and decreases phagocytosis of C3bi-opsonized and non-opsonized myelin by primary mouse microglia.