Anti-Nuf2 antibody, rabbit serum

Description

Nuf2 is a component of kinetochore-associated Ndc80 complex, which serves to attach microtubules to the kinetochore.Nuf2 is a conserved protein from yeast to human. Disruption of the Nuf2 gene in Schizosaccharomyces pombe causes defect in chromosome segregation and in the spindle checkpoint.  Yeast Nuf2 disappears from the centromere during meiotic prophase when centromeres lose their connection to the spindle pole body, and plays a regulatory role in chromosome segregation. In human cells, Nuf2 specifically functions at kinetochores for stable microtubule attachment. Down regulation of the protein by RNA interference results in failure of the kinetochores to form attachments to the spindle microtubules. As a result, cells are blocked in the prometaphase stage with an active spindle checkpoint and undergo cell death. This antibody was prepared and tested by Prof. Tokuko Haraguchi at Kobe Advanced ICT Research Center, National Institute of Information and Communications Technology

Applications

1. Western blot 500~5,000 fold dilution
2. Immunofluorescence staining 200~1,000 fold dilution

Methanol fixation is recommended for immunofluorescence staining.

Specification

Immunogen: Synthetic peptide CGGDSYAKIDEKTAELKRKMFKMS corresponding to the C-terminus region of human Nuf2

Specificity: React with human and chicken Nuf2. Not tested with other species.

Form: Whole serum added with 0.05% sodium azide

Storage: Shipped at 4°C and long term storage at -20°C

Data Link

UniProtKB/Swiss-Prot　Q9BZD4 (NUF2_HUMAN)

References:

1. Wigge PA and Kilmaetin JV “The Ndc80p complex from Saccharomyces cerevisiae contains conserved centromere components and has a function in chromosome segregation.” J Cell Biol 152: 349-360 (2001) PMID: 11266451
2. Nabetani A, Koujin T, Tsutsumi C, Haraguchi T, Hiraoka Y “A conserved protein, Nuf2, is implicated in connecting the centromere to the spindle during chromosome segregation: a link between the kinetochore function and the spindle checkpoint.”Chromosoma 110: 322-334 (2001) PMID: 11685532
3. DeLuca JG, Moree B, Hickey JM, Kilmartin JV, Salmon ED “hNuf2 inhibition blocks stable kinetochore-microtubule attachment and induces mitotic cell death in HeLa cells.” J Cell Biol 159: 549-555 (2002) PMID: 12438418

Fig. 1 Detection of Nuf2 protein in HeLa cells by Western blot using this antibody.

The antibody was diluted to 5,000 fold.

Human Nuf2 protein was detected as a 50 kD band (predicted mass is 54 kD).

Fig. 2 Detection of Nuf2 protein in HeLa cells by immunofluorescence staining using this antibody. HeLa cells were fixed with methanol and subjected to immunofluorescence staining using this antibody. The antibody was diluted to 500 fold. Chromosomes were stained by DAPI (violet). Nuf2 protein (green) actively localizes to kinetochores at the stages of prometaphase to anaphase.