Methods and compositions for modulating axonal outgrowth of central nervous system neurons
a central nervous system and axonal outgrowth technology, applied in the direction of depsipeptides, peptide/protein ingredients, powder delivery, etc., can solve the problems of inability to re-grow severed connections, cognitive, sensory and motor function loss, and loss of life-long vital functions
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Isolation and Characterization of the N-kinase Polypeptide
[0116] Neocortical gray matter from bovine brain was homogenized in buffer containing protease and phosphatase inhibitors, and particulate material was centrifuged down. The soluble fraction from approximately 1 kg of tissue was used as the starting material for isolation of the kinase.
[0117] During the purification process, kinase activity was monitored using an in-gel kinase method. In this method, a histone HF-1 substrate protein is polymerized into a 10% polyacrylamide gel before the samples to be analyzed are electrophoresed in the gel. Following completion of the electrophoresis, proteins are partially renatured with guanidium isothiocyanate and incubated in the presence [32P]-ATP plus Mn2+, with or without 6-TG present. (Activation by Mn2+, but not by Mg2+ or Ca2+, is a distinctive property of N-kinase). The purification of the kinase was monitored by looking for a 6-TG-inhibitable radioactive band corresponding to t...
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