K0171-3 For Research Use Only. Page 1 of 2 Not for use in diagnostic procedures.

MONOCLONAL ANTIBODY Anti-CENP-E Code No. Clone Subclass Quantity Concentration K0171-3 1H12 Mouse IgG1 100 g 1 mg/mL

BACKGROUND: -E (CENP-E) is a INTENDED USE: -like protein that binds to during For Research Use Only. Not for use in diagnostic procedures. . CENP-E is required for full alignment and normal progression through mitosis. REFERENCES: CENP-E is essential for stable, bioriented attachment of 1) Liu, S. T., et al., Mol. Biol. Cell 14, 1638-1651 (2003) to spindle microtubules, for development of 2) McEwen, B. F., et al., Mol. Biol. Cell 12, 2776-2789 (2001) tension across aligned chromosomes, for stabilization of 3) Sudakin, V., et al., J. Cell Biol. 154, 925-936 (2001) spindle poles, and for satisfying the mitotic checkpoint. 4) Chan, G. K. T., et al., J. Cell Biol. 146, 941-954 (1999) The localization of CENP-E differs markedly from other centromere , CENP-A, CENP-B, CENP-C and CENP-D. In contrast to these antigens, CENP-E RELATED PRODUCTS: expression is appeared at the centromere region of D115-3 anti-CENP-A (3-19) chromosomes during prometaphase but is not detected PD018 anti-CENP-K (polyclonal) during . PD019 anti-CENP-50 (polyclonal) PD020 anti-CENP-O (polyclonal)

SOURCE: This antibody was purified from hybridoma (clone 1H12) supernatant using protein A agarose. This hybridoma was established by fusion of mouse myeloma cell Sp2/0 with Balb/c mouse splenocyte immunized with the recombinant full-length CENP-E.

FORMULATION: 100 g IgG in 100 L volume of PBS containing 50% glycerol, pH 7.2. No preservative is contained.

STORAGE: This antibody solution is stable for one year from the date of purchase when stored at -20°C.

REACTIVITY: This antibody reacts with CENP-E on Immunocytochemical detection of Immunocytochemistry. CENP-E on acetone fixed HEp-II cells with K0171-3. APPLICATIONS: Western blotting; Not recommended PROTOCOL: Immunoprecipitation; Not tested Immunocytochemistry Immunohistochemistry; Not recommended 1) Culture the cells in the appropriate condition on a glass Immunocytochemistry; 10 g/mL slide. (for example, spread 104 of cells per one well, then Flow cytometry; Not tested incubate in a CO2 incubator for one night.) 2) Fix the cells by immersing the slide in acetone for 10 Detailed procedure is provided in the following minutes on ice. PROTOCOL. 3) Air dry the slides. 4) Add the primary antibody diluted with PBS as suggest in the APPLICATIONS onto the cells and incubate for 1 SPECIES CROSS REACTIVITY: hour at room temperature. (Optimization of antibody Species Human Mouse Rat concentration or incubation condition are recommended Cell HEp-II Not Tested Not Tested if necessary.) 5) Prepare a wash container such as a 500 mL beaker with a Reactivity on IF + magnetic stirrer. Then wash the cultured cells on the glass slide by soaking the slide with a plenty of PBS in

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the wash container for 5 minutes. Take care not to touch the cells. Repeat another washes once more. 6) Add 30 L of 1:100 FITC conjugated anti-mouse IgG (MBL; code no. 238) diluted with PBS onto the cells. Incubate for 30 minutes at room temperature. Keep out light by aluminum foil. 7) Wash the slide in a plenty of PBS as in the step 5). 8) Wipe excess liquid from slide but take care not to touch the cells. Never leave the cells to dry. 9) Promptly add PermafluorTM aqueous mounting medium (MBL; code no. IM-0752) onto the slide, then put a cover slip on it.

(Positive control for Immunocytochemistry; HEp-II)

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