1/500 - 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Promotes ATP-dependent removal of tubulin dimers from microtubules. Regulates the turnover of microtubules at the kinetochore and functions in chromosome segregation during mitosis.
Expressed at high levels in thymus and testis, at low levels in small intestine, the mucosal lining of colon, and placenta, and at very low levels in spleen and ovary; expression is not detected in prostate, peripheral blood Leukocytes, heart, brain, lung, liver, skeletal muscle, kidney or pancreas. Isoform 2 is testis-specific.
Belongs to the kinesin-like protein family. MCAK/KIF2 subfamily. Contains 1 kinesin-motor domain.
Isoform 2 is expressed in fetal testis.
The microtubule tip localization signal (MtLS) motif; mediates interaction with MAPRE1 and targeting to the growing microtubule plus ends.
Phosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation by STK12, regulates association with centromeres and kinetochores and the microtubule depolymerization activity.
Cytoplasm > cytoskeleton. Nucleus. Chromosome > centromere. Chromosome > centromere > kinetochore. Associates with the microtubule network at the growing distal tip (the plus-end) of microtubules, probably through interaction with MTUS2/TIP150 and MAPRE1 (By similarity). Centromeric localization requires the presence of BUB1 and SGOL2.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse hybridoma tumor tissue labelling MCAK with ab71706 at 1/1000 (1µg/ml). Detection: DAB.
Western blot - MCAK antibody (ab71706)
All lanes : Anti-MCAK antibody (ab71706) at 0.1 µg/ml
Lane 1 : NIH3T3 whole cell lysate at 50 µg/ml Lane 2 : NIH3T3 whole cell lysate at 15 µg/ml Lane 3 : NIH3T3 whole cell lysate at 5 µg/ml
Predicted band size: 81 kDa
Exposure time: 3 minutes
Immunoprecipitation - MCAK antibody (ab71706)
MCAK antibody (ab71706) recognizing MCAK protein in NIH3T3 whole cell lysate (1 mg, 20% of IP loaded).
Lane 1: MCAK antibody (ab71706) used for IP at 3 µg/mg lysate and for WB at 1.0 µg/ml.
Lane 2: Non-Abcam MCAK antibody, specific for a downstream epitope of MCAK.
Lane 3: Control IgG.
Detection by chemiluminescence with exposure time of 30 seconds.