Anti-INCENP Developed in Rabbit IgG Fraction of Antiserum Product Number I 5283 Product Description Anti-INCENP is developed in rabbit using as Reagent immunogen a synthetic peptide corresponding to amino Anti-INCENP is supplied as a solution in acid 884-901 of human INCENP with N-terminal added 0.01 M phosphate buffered saline, pH 7.4, containing cysteine, conjugated to KLH. The corresponding 0.02 15 mM sodium azide. sequence differs by one amino acid in chicken and Xenopus laevis and by two amino acids in mouse. Precautions and Disclaimer Whole antiserum is fractionated and then further Due to the sodium azide content, a material safety data purified by ion-exchange chromatography to provide the sheet (MSDS) for this product has been sent to the IgG fraction of antiserum that is essentially free of other attention of the safety officer of your institution. Consult rabbit serum proteins. the MSDS for information regarding hazards and safe handling practices. Anti-INCENP recognizes human, mouse, and rat INCENP. Applications include the detection of INCENP Storage/Stability by immunoblotting (~106 kDa) and For continuous use, store at 2-8 °C for up to one month. immunofluorescence. Detection of the INCENP band by For prolonged storage, freeze in working aliquots at immunoblotting is specifically inhibited with the −20 °C. Repeated freezing and thawing is not immunizing peptide. recommended. Storage in frost-free freezers is also not recommended. If slight turbidity occurs upon prolonged Chromosomal passenger proteins are members of a storage, clarify the solution by centrifugation before group of proteins that move from centromeres to the use. Working dilutions should be discarded if not used 1 spindle midzone during mitosis. INCENP (Inner within 12 hours. Centromere Protein) is a chromosomal passenger 1-6 protein with an essential role in mitosis and meiosis. Product Profile It is a complex highly basic multidomain protein that is For immunoblotting, a minimum working antibody dynamically translocated from metaphase dilution of 1:5,000 is recommended using a nuclear chromosomes to the spindle midzone during or just extract of human HeLa cells and a chemiluminescent 3 prior to anaphase. Some INCENP transfers to the detection reagent. equatorial cortex before formation of the cleavage 4 furrow. Ultimately, it is discarded in the midbody at the For immunoblotting, a minimum working antibody 5 completion of cytokinesis. INCENP, in complex with dilution of 1:2,000 is recommended using a whole the passenger proteins aurora-B kinase and survivin, extract of mouse NIH-3T3 and rat PC12 cells and a 6 acts at multiple points during mitosis. Studies in chemiluminescent detection reagent. knockout mice established INCENP as essential for mouse development and viability. Loss of INCENP For indirect immunofluorescence, a minimum working results in chromosome segregation defects and failures antibody dilution of 1:100 is recommended using 5 in cytokinesis. Targeting of survivin to its centromeric human HeLa cells. and central spindle locations during mitosis is 6 dependent on INCENP. Direct binding of INCENP to Note: In order to obtain the best results using various β-tubulin has been demonstrated.4 Interactions of techniques and preparations, we recommend deter- α INCENP with heterochromatin protein HP1Hsa and mining the optimal working dilutions by titration. histone H2A.Z were also reported.7, 8 INCENP levels are increased in several human cancer lines.3 References 5. Cutts, S., et al., Hum. Mol. Genet., 8, 1145-1155 1. Cooke, C.A., et al., J. Cell Biol., 105, 2053-2067 (1999). (1987). 6. Wheatley, S.P., et al., Curr.Biol., 11, 886-890 2. Parra, M.T., et al., J. Cell Sci., 116, 961-974 (2003). (2001). 3. Adams, R.R., et al., Chromosoma, 110, 65-74 7. Ainsztein, A.M., et al., J. Cell Biol., 143, 1763-1774 (2001). (1998). 4. Wheatley, S.P., et al., Exp. Cell Res., 262, 122-127 8. Rangsamy, D., et al., EMBO J., 22, 1599-1607 (2001). (2003). ST/KAA 11/03 Sigma brand products are sold through Sigma-Aldrich, Inc. Sigma-Aldrich, Inc. warrants that its products conform to the information contained in this and other Sigma-Aldrich publications. Purchaser must determine the suitability of the product(s) for their particular use. Additional terms and conditions may apply. Please see reverse side of the invoice or packing slip. .