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 210 kDa (predicted molecular weight: 186 kDa).
Use at 2-5 µg/mg of lysate.
1/200 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
1/200 - 1/1000.
Permeabilization with Triton-X 100 is recommended for formaldehyde-fixed cells.
Use at an assay dependent concentration.
FunctionPlays an essential role in cytokinesis.
Sequence similaritiesBelongs to the kinesin-like protein family. Contains 1 FHA domain. Contains 1 kinesin-motor domain.
Developmental stageUp-regulated in cells progressing through G2/M phase.
Cellular localizationNucleus. Cytoplasm. Cytoplasm > cytoskeleton > spindle. Nuclear localization observed during interphase in PubMed:16431929 or triggered by entry into mitosis in PubMed:16648480. Cytoplasmic in interphase (PubMed:16648480) and metaphase cells (PubMed:16431929). From prophase to metaphase, accumulates at the developing spindle poles and their associated microtubules. During anaphase, accumulates at the spindle midzone. Localization to the central spindle and midbody during anaphase is dependent upon PRC1 and CIT presence. In cells ready to undergo abscission, concentrates at the contractile ring.
Immunocytochemistry/Immunofluorescence analysis of formaldelyde-fixed HeLa cells labelling KIF14 with ab71155 at 1/100 (2µg/ml). A red-fluorescent goat anti-rabbit IgG (1/100) was used as the secondary antibody.
Western blot - KIF14 antibody (ab71155)
All lanes : Anti-KIF14 antibody (ab71155) at 0.04 µg/ml
Lane 1 : Whole cell lysate from Hela cells at 50 µg Lane 2 : Whole cell lysate from Hela cells at 15 µg Lane 3 : Whole cell lysate from Hela cells at 5 µg Lane 4 : Whole cell lysate from 293T cells at 50 µg