The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 4 µg/ml. The following steps are recommended prior to antibody incubation: cells fixation in ice cold 4% paraformaldehyde pH 7.2-7.3, followed by permeabilization with 0.1% Triton X-100 in PBS for 3x5 minutes.
1/250 - 1/500.
1/200 - 1/500. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
FunctionMotor protein that translocates PRC1 to the plus ends of interdigitating spindle microtubules during the metaphase to anaphase transition, an essential step for the formation of an organized central spindle midzone and midbody and for successful cytokinesis. May play a role in mitotic chromosomal positioning and bipolar spindle stabilization.
Tissue specificityHighly expressed in hematopoietic tissues, fetal liver, spleen, thymus and adult thymus and bone marrow. Lower levels are found in heart, testis, kidney, colon and lung.
Sequence similaritiesBelongs to the kinesin-like protein family. Chromokinesin subfamily. Contains 1 kinesin-motor domain.
Cellular localizationNucleus matrix. Cytoplasm > cytoskeleton > spindle. Midbody. Chromosome. Not present in the nucleolus. In early mitosis, associated with the mitotic spindle, in anaphase, localized to the spindle midzone and, in telophase and cytokinesis, to the midbody. In late cytokinesis, found in the center of the midbody. Associated with chromosomes at all stages of mitosis.