Intracellular Staining by Flow Cytometry
|Detection of BDNF in U‑87 MG Human Cell Line by Flow Cytometry. U‑87 MG human glioblastoma/astrocytoma cell line was stained with Mouse Anti-Human BDNF PerCP‑conjugated Monoclonal Antibody (Catalog # IC2481C, filled histogram) or isotype control antibody (Catalog # IC002C, open histogram). To facilitate intracellular staining, cells were fixed with Flow Cytometry Fixation Buffer (Catalog # FC004) and permeabilized with Flow Cytometry Permeabilization/Wash Buffer I (Catalog # FC005). View our protocol for Staining Intracellular Molecules.|
Brain-derived neurotrophic factor (BDNF) is a member of the NGF family of neurotrophic factors (also named neurotrophins) that are required for the differentiation and survival of specific neuronal subpopulations in both the central as well as the peripheral nervous system. The neurotrophin family is comprised of at least four proteins including NGF, BDNF, NT-3, and NT-4/5. These secreted cytokines are synthesized as prepropeptides that are proteolytically processed to generate the mature proteins. All neurotrophins have six conserved cysteine residues that are involved in the formation of three disulfide bonds and all share approximately 55% sequence identity at the amino acid level. Similarly to NGF, bioactive BDNF is predicted to be a non-covalently linked homodimer.
BDNF cDNA encodes a 247 amino acid precursor protein with a signal peptide and a proprotein that are cleaved to yield the 119 amino acid mature BDNF. The amino acid sequence of mature BDNF is identical in all mammals examined. High levels of expression of BDNF have been detected in the hippocampus, cerebellum, fetal eye, and placenta. In addition, low levels of BDNF expression are found in the pituitary gland, spinal cord, heart, lung, and skeletal muscle. BDNF binds with high affinity and specifically activates the TrkB tyrosine kinase receptor.