SpecificityThis antibody recognizes a protein of approximately 31kDa in HeLa cell lysate that is blocked by the addition of the immunizing peptide. This is larger than the expected size of 25kDa and we cannot explain why the detected protein is larger than expected. Several larger bands are detected which are not blocked at all by the addition of the immunizing peptide. SPC25 may be better detected in lysates of nocodazole-arrested HeLa cells.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application notesICC/IF: 1/200.
WB: Use at a concentration of 1 µg/ml. Detects a band of approximately 28 kDa (predicted molecular weight: 26 kDa according to SwissProt data for mouse SPC25).
Not yet tested in other applications.
Optimal dilutions/concentrations should be determined by the end user.
FunctionActs as a component of the essential kinetochore-associated NDC80 complex, which is required for chromosome segregation and spindle checkpoint activity. Required for kinetochore integrity and the organization of stable microtubule binding sites in the outer plate of the kinetochore.
Sequence similaritiesBelongs to the SPC25 family.
Cellular localizationNucleus. Chromosome > centromere > kinetochore. Localizes to kinetochores from late prophase to anaphase. Localizes specifically to the outer plate of the kinetochore.
Immunocytochemistry/ Immunofluorescence - SPC25 antibody (ab20679)This image is courtesy of an Abreview submitted by Dr Kirk McManus
ab20679 (1/200) staining SPC25 in assynchronous HeLa cells, in conjunction with a secondary goat anti-rabbit antibody conjugated to Cy3® (green). Cells were counterstained with DAPI to show the nucleus (red).