The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 µg/ml. Predicted molecular weight: 34 kDa. Good results were obtained when blocked with 5% non-fat dry milk in 0.05% PBS-T.
Functionp35 is a neuron specific activator of CDK5. The complex p35/CDK5 is required for neurite outgrowth and cortical lamination. Activator of TPKII.
Tissue specificityBrain and neuron specific.
Sequence similaritiesBelongs to the cyclin-dependent kinase 5 activator family.
Post-translational modificationsThe p35 form is proteolytically cleaved by calpain, giving rise to the p25 form. P35 has a 5 to 10 fold shorter half-life compared to p25. The conversion results in deregulation of the CDK5 kinase: p25/CDK5 kinase displays an increased and altered tau phosphorylation in comparison to the p35/CDK5 kinase in vivo. Probably myristoylated. The Gly-2-Ala mutant is absent of the cell periphery, suggesting that a proper myristoylation signal is essential for the proper distribution of p35.
Cellular localizationCell membrane. In the primary cortical neurons, p35 is present in the peripheries and nerve terminals and Nucleus. Cytoplasm > perinuclear region. The conversion of p35 to p25 relocalizes the protein from the cell periphery to the cytoplasm, in nuclear and perinuclear regions. In the primary cortical neurons, p25 is primarily concentrated in the cell soma and is largely absent from neurites.