Anti-BUB3 produced in rabbit, affinity isolated antibody

Catalog Number B7811

Product Description Precautions and Disclaimer Anti-BUB3 is produced in rabbit using as immunogen a This product is for R&D use only, not for drug, synthetic peptide corresponding to amino acids 307-325 household, or other uses. Please consult the Material of human BUB3 ( ID: 9184), conjugated to KLH. Safety Data Sheet for information regarding hazards This sequence differs in mouse and rat by two and and safe handling practices. three amino acids, respectively. The antibody is affinity purified using the immunizing peptide immobilized on Storage/Stability agarose. For continuous use, store at 2-8 °C for up to one month. For extended storage, freeze in working aliquots. Anti-BUB3 specifically recognizes BUB3 (37 kDa). Repeated freezing and thawing, or storage in “frost- Applications include immunoblotting, immuno- free” freezers, is not recommended. If slight turbidity precipitation, and immunofluorescence. Detection of the occurs upon prolonged storage, clarify the solution by BUB3 band in immunoblotting is specifically inhibited by centrifugation before use. Working dilutions should be the immunizing peptide. discarded if not used within 12 hours.

Eukaryotes developed a feedback control mechanism, Product Profile also referred as a cell cycle checkpoint, to ensure Immunoblotting: a working concentration of 2-4 mg/mL accurate segregation during mitosis, is recommended using NIH-3T3 and Rat1 total cell which delays metaphase- transition until all lysates. the are correctly aligned on the 1 microtubule spindle apparatus. The first evidence Immunoprecipitation: a working amount of 3-6 mg is demonstrating that onset of anaphase is regulated by a recommended using HEK 293-T cell lysates. cell cycle checkpoint came from c.elegance where genetic screens have identified a series of Immunofluorescence: a working concentration of (BUB1, BUB2, BUB3, , , MAD3, and MPS1) 2, 3, 4 1-2 mg/mL is recommended using paraformaldehyde that fail to arrest in response to spindle damage as fixed HEK293-T cells. necessary for the preanaphase delay. Eukaryote orthologs of these genes are Bub1, Bub3, Note: In order to obtain the best results using various BubR1/Mad3, Mad1, and Mad2. These were techniques and preparations, we recommend deter- further shown to localize preferentially to unattached mining the optimal working dilutions by titration. .5 Bub3 contains four conserved WD40 repeats that are known to be important for - References 6 protein interactions. Genetic disruption of the Bub3 1. Hartwell, L.H., and Weinert, T.A., Science, 246, gene in mouse revealed a lethal phenotype for the 629–634 (1989). gene-disrupted mice and show that Bub3 is an essential 2. Hoyt, M.A., et al., Cell, 66, 507-517 (1991). component of the mitotic spindle checkpoint pathway 3. Li, R., and Murray, A.W., Cell, 66, 519-531 (1991). 7 that is required for mitotic fidelity. 4. Weiss, E., and Winey, M.J., Cell. Biol., 132, 111- 123 (1996). Reagent 5. Lopes, C.S., et al., J. Cell Sci. 118, 187-198 Supplied as a solution in 0.01 M phosphate buffered (2005). saline, pH 7.4, containing 15 mM sodium azide as a 6. Neer, E.J., et al., Nature, 371, 297-300 (1994). preservative. 7. Kalitsis, P., at al., Gene & Development 14, 2277- 2282 (2000). Antibody concentration: 1 mg/mL SG,CS,PHC 01/08-1

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