ab60006 is first purified by protein G affinity chromatography. Then, the antibody fraction is peptide affinity purified in a 2 step procedure with the control and phosphorylated peptides. The phospho specific antibody is eluted with high and low pH buffers and neutralized immediately, followed by dialysis against PBS.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use at an assay dependent concentration. PubMed: 24785679
1/50 - 1/100.
Required for organization of the cellular microtubule array and microtubule anchoring at the centrosome. May regulate microtubule organization at least in part by targeting the microtubule severing protein KATNA1 to the centrosome. Also positively regulates the activity of the minus-end directed microtubule motor protein dynein. May enhance dynein-mediated microtubule sliding by targeting dynein to the microtubule plus ends. Required for several dynein- and microtubule-dependent processes such as the maintenance of Golgi integrity, the centripetal motion of secretory vesicles and the coupling of the nucleus and centrosome. Also required during brain development for the migration of newly formed neurons from the ventricular/subventricular zone toward the cortical plate. Plays a role, together with DISC1, in the regulation of neurite outgrowth. Required for mitosis in some cell types but appears to be dispensible for mitosis in cortical neuronal progenitors, which instead requires NDE1. Facilitates the polymerization of neurofilaments from the individual subunits NEFH and NEFL.
Expressed in brain, heart, kidney, liver, lung, pancreas, placenta and skeletal muscle.
Belongs to the nudE family.
Expression peaks in mitosis.
Phosphorylated in mitosis. Can be phosphorylated by CDK1, CDK5 and MAPK1. Phosphorylation by CDK5 promotes interaction with KATNA1 and YWHAE.
Cytoplasm > cytoskeleton. Cytoplasm > cytoskeleton > centrosome. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > spindle. Localizes to the cell body of the motor neurons and colocalizes with assembled neurofilaments within axonal processes. Localizes to the microtubules of the manchette in elongated spermatids. Colocalizes with DISC1 in the perinuclear region, including the centrosome (By similarity). Localizes to the interphase centrosome and the mitotic spindle. Localizes to the kinetochore in a CENPF-dependent manner.
ab60006 at 1/50 dilution staining Nudel in Human breast carcinoma by Immunohistochemistry, Formalin-fixed, Paraffin-embedded tissue.
The primary antibody was peroxidase conjugated to the secondary antibody, followed by AEC staining.