The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
1/2000 - 1/10000. Detects a band of approximately 120 kDa (predicted molecular weight: 120 kDa).
Use at 2-5 µg/mg of lysate.
Motor protein required for establishing a bipolar spindle. Blocking of KIF11 prevents centrosome migration and arrest cells in mitosis with monoastral microtubule arrays.
Involvement in disease
Defects in KIF11 are the cause of microcephaly with or without chorioretinopathy, lymphedema, or mental retardation (MCLMR) [MIM:152950]. An autosomal dominant disorder that involves an overlapping but variable spectrum of central nervous system and ocular developmental anomalies. Microcephaly ranges from mild to severe and is often associated with mild to moderate developmental delay and a characteristic facial phenotype with upslanting palpebral fissures, broad nose with rounded tip, long philtrum with thin upper lip, prominent chin, and prominent ears. Chorioretinopathy is the most common eye abnormality, but retinal folds, microphthalmia, and myopic and hypermetropic astigmatism have also been reported, and some individuals have no overt ocular phenotype. Congenital lymphedema, when present, is typically confined to the dorsa of the feet, and lymphoscintigraphy reveals the absence of radioactive isotope uptake from the webspaces between the toes.
Belongs to the kinesin-like protein family. BimC subfamily. Contains 1 kinesin-motor domain.
Phosphorylated exclusively on serine during S phase, but on both serine and Thr-926 during mitosis, so controlling the association of KIF11 with the spindle apparatus (probably during early prophase). Phosphorylated upon DNA damage, probably by ATM or ATR. A subset of this protein primarily localized at the spindle pole is phosphorylated by NEK6 during mitosis; phosphorylation is required for mitotic function.
Cytoplasm. Cytoplasm > cytoskeleton > spindle pole.
All lanes : Anti-Eg5 antibody (ab72413) at 0.04 µg/ml
Lane 1 : HeLa whole cell lysate at 50 µg Lane 2 : HeLa whole cell lysate at 15 µg Lane 3 : HeLa whole cell lysate at 5 µg Lane 4 : 293T whole cell lysate at 50 µg
Predicted band size: 120 kDa Observed band size: 120 kDa
Exposure time: 3 seconds
Immunoprecipitation - Anti-Eg5 antibody (ab72413)
Detection of Eg5 by Western Blot of Immunprecipitate.
ab72413 at 1µg/ml staining Eg5 in HeLa whole cell lysate immunoprecipitated using ab72413 at 3µg/mg lysate (1 mg/IP; 20% of IP loaded). Detection: Chemiluminescence with exposure time of 1 second.