Microtubule plus-end tracking protein that promotes the stabilization of dynamic microtubules. Involved in the nucleation of noncentrosomal microtubules originating from the trans-Golgi network (TGN). Required for the polarization of the cytoplasmic microtubule arrays in migrating cells towards the leading edge of the cell. May act at the cell cortex to enhance the frequency of rescue of depolymerizing microtubules by attaching their plus-ends to cortical platforms composed of ERC1 and PHLDB2. This cortical microtubule stabilizing activity is regulated at least in part by phosphatidylinositol 3-kinase signaling. Also performs a similar stabilizing function at the kinetochore which is essential for the bipolar alignment of chromosomes on the mitotic spindle.
Belongs to the CLASP family. Contains 7 HEAT repeats.
Phosphorylated upon DNA damage, probably by ATM or ATR.
Cytoplasm > cytoskeleton. Cytoplasm > cytoskeleton > centrosome. Chromosome > centromere > kinetochore. Cytoplasm > cytoskeleton > spindle. Golgi apparatus > trans-Golgi network. Localizes to microtubule plus ends. Localizes to centrosomes, kinetochores and the mitotic spindle from prometaphase. Subsequently localizes to the spindle midzone from anaphase and to the midbody from telophase. In migrating cells localizes to the plus ends of microtubules within the cell body and to the entire microtubule lattice within the lamella. Localizes to the cell cortex and this requires ERC1 and PHLDB2.
Western blot - Anti-CLASP1 antibody [KT67] (ab95372)
All lanes : Anti-CLASP1 antibody [KT67] (ab95372) at 1/1000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate Lane 2 : CLASP1 knockout HAP1 whole cell lysate Lane 3 : HeLa whole cell lysate Lane 4 : Jurkat whole cell lysate
Lysates/proteins at 20 µg per lane.
Predicted band size: 169 kDa
Lanes 1 - 4: Merged signal (red and green). Green - ab95372 observed at 169 kDa. Red - loading control, ab176560, observed at 50 kDa.
ab95372 was shown to specifically react with CLASP1 in wild-type HAP1 cells as signal was lost in CLASP1 knockout cells. Wild-type and CLASP1 knockout samples were subjected to SDS-PAGE. Ab95372 and ab176560 (Rabbit anti-alpha Tubulin loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rat IgG H&L (IRDye® 800CW) and Goat anti-Rabbit IgG H&L (IRDye® 680RD) preabsorbed ab216777 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.