This antibody gave a signal in the following whole cell lysates;
HeLa (Human epithelial carcinoma cell line)
Jurkat (Human T cell lymphoblast-like cell line)
A431 (Human epithelial carcinoma cell line)
HEK293 (Human embryonic kidney cell line)
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C or -80°C. Avoid freeze / thaw cycle.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesIHC-P: Use at a concentration of 5 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF: Use at a concentration of 1 µg/ml.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 75 kDa (predicted molecular weight: 81 kDa).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionPromotes ATP-dependent removal of tubulin dimers from microtubules. Regulates the turnover of microtubules at the kinetochore and functions in chromosome segregation during mitosis.
Tissue specificityExpressed 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.
Sequence similaritiesBelongs to the kinesin-like protein family. MCAK/KIF2 subfamily. Contains 1 kinesin-motor domain.
Developmental stageIsoform 2 is expressed in fetal testis.
DomainThe microtubule tip localization signal (MtLS) motif; mediates interaction with MAPRE1 and targeting to the growing microtubule plus ends.
Post-translational modificationsPhosphorylated upon DNA damage, probably by ATM or ATR. Phosphorylation by STK12, regulates association with centromeres and kinetochores and the microtubule depolymerization activity.
Cellular localizationCytoplasm > 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.
ICC/IF image of ab42676 stained human HeLa cells. The cells were PFA fixed (10 min), permabilised in PBS-T (20 min) and incubated with the antibody (ab42676, 1µg/ml) for 1h at room temperature. 1%BSA / 10% normal goat serum / 0.3M glycine was used to quench autofluorescence and block non-specific protein-protein interactions. The secondary antibody (green) was Alexa Fluor® 488 goat anti-rabbit IgG (H+L) used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red). DAPI was used to stain the cell nuclei (blue).
IHC image of MCAK staining in human liver carcinoma FFPE section, performed on a BondTM system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab42676, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
References for Anti-MCAK antibody (ab42676)
has not yet been referenced specifically in any publications.
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