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Centromere Protein E, 312kDa (CENPE) antibody

Details for Product No. ABIN112051
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Antigen
Synonyms CENPE, kif10, cenp-E, XCENP-E, 312kDa, AU019344, BC049989, C530022J18, CENP-E, Kif10, KIF10, PPP1R61
Reactivity
Human
(23), (2), (1), (1)
Host
Mouse
(14), (8), (3)
Clonality (Clone)
Monoclonal ()
Conjugate
Un-conjugated
(1), (1), (1), (1), (1), (1)
Application
Immunocytochemistry (ICC), Immunofluorescence (IF), Immunoprecipitation (IP), Western Blotting (WB)
(19), (10), (10), (7), (6), (2), (2), (2)
Pubmed 3 references available
Quantity 0.1 mL
Shipping to United States (Change)
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Catalog No. ABIN112051
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Immunogen Recombinant human CENP-E.
Clone 1H12
Isotype IgG1
Specificity Human CENP-E. Not tested on reactivity with rodents or other vertebrates. Species Reactivity: Tested: Human.
Purification Ion exchange chromatography.
Alternative Name CENPE
Background CENPE is a 250-300 kDa human centromere-associated kinesin-like motor protein that accumulates in G2 phase. In contrast to other centromere proteins, CENPE is not detected at centromeres during interphase, and first appears at the centromere region of chromosomes during prometaphase. CENPE function is required for the transition from metaphase to anaphase. CENPE is probably one of the motors responsible for mammalian chromosome movement and/or spindle elongation.
Alternate names: CENP-E, Centromere-associated protein E, Centromeric protein E, Kinesin-related protein CENPE
Gene ID 1062
NCBI Accession NP_001804.2
UniProt Q02224
Research Area Chromatin and Nuclear Signaling, Cell Cycle
Application Notes Immunoprecipitation: Use ~3 mg/ml of antibody for 300 µg of cell lysate. Immunocytochemistry: Use 0.5-1 µg/ml as a guideline. Western blot: The antigen is first concentrated by Immunoprecipitation. Any Human cell line should be used for a Positive Control. Other applications not tested. Optimal dilutions are dependent on conditions and should be determined by the user.
Protocol Immunofluorescence protocol - Formaldehyde fixationCollect cells from T. c. unit and remove media from petri dish using suction. Wash with 1x PBS and remove. Incubate cells in pre-warm (37°C) Para-Formaldehyde for 12 minutes at room temperatureon an orbital shaker. Remove PFA and incubate in 0. 5% Triton X-IOO in 1x PBS for 5 minutes at roomtemperature. Prepare blocking reagent, this is also the antibody diluent. Wash cells 2x with 1x PBS at room temperature, for 4 minutes/wash on an orbital shaker. Block with 1 % NCS and 1x PBS for 30 minutes at room temperature. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. Wash cells 2x with lx PBS at room temperature and air dry briefly. Incubate with primary antibody for 1 hr at room temperature in the dark in stainingchambers. During this time prepare the secondary antibody. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)Incubate with secondary antibody for 1 hour at room temperature in the dark in stainingchambers. Wash cells 5x with 1x PBS. Mount in Dapi. Solutions (prepare fresh the same day of staining). 1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh l0x PBS). Fixation solution: 3. 5% Para formaldehyde. 1. 75g PFA in 20 ml d. H20 plus 5 drops 1M NaOH. Stir on a hot plate at 50-60°C untildissolved. Add 4 drops IN HCI and check pH indicator strip. PH should be 7. 4. Completevolume with d. H20 to 25ml and add 25ml 2xPBS. Check pH before adding to cover slips. Immunofluorescence protocol - Methanol/acetone fixationCollect cells from T. C. unit and remove media from petri dish using suction. Wash with 1x PBS and remove. Fix cells with cold methanol: acetone 1: 1 for 10 minutes on ice. Prepare blocking reagent, this is also the diluent for the antibodies. Remove fixative and wash cells 3x with Ix PBS at RT, for 4 minutes/wash on orbital shaker. Block with 1% NCS and Ix PBS for 30 minutes at RT. Prepare primary antibodies (50μl/coverslip) and moist staining chambers. Wash cells 2x with 1 x PBS at RT and air dry for approximately 7 minutes. Incubate with primary antibody for 1 hr at RT in the dark in staining chambers. During thistime prepare secondary antibody. Wash cells 5x with 1x PBS (5 beaker changes/5 counts in each beaker)Incubate with secondary antibody for 1 hr at R T in the dark in staining chambers. Wash cells 5x with 1x PBS. Mount in Dapi. Solutions (prepare fresh the same day of staining)1x Phosphate buffered saline. Blocking reagent: 1% NCS in 1x PBS (use fresh 10x PBS). Fixation solution: methanol: acetone 1: 1 ice cold. Western Blotting ProtocolTransfer gel to PDVF or nitrocellulose membranePlace membrane in plastic tray in blocking buffer for one hour with agitation
Restrictions For Research Use only
Format Liquid
Concentration 0.45 mg/mL
Buffer PBS with 0.09% sodium azide as preservative.
Preservative Sodium azide
Precaution of Use This product contains sodium azide: a POISONOUS AND HAZARDOUS SUBSTANCE which should be handled by trained staff only.
Handling Advice Avoid repeated freezing and thawing.
Storage 4 °C/-20 °C
Storage Comment Store the antibody at 2-8°C for one month or (in aliquots) at -20°C for longer.
Expiry Date 12 months
Background publications Yen, Li, Schaar et al.: "CENP-E is a putative kinetochore motor that accumulates just before mitosis." in: Nature, Vol. 359, Issue 6395, pp. 536-9, 1992 (PubMed).

Yen, Compton, Wise et al.: "CENP-E, a novel human centromere-associated protein required for progression from metaphase to anaphase." in: The EMBO journal, Vol. 10, Issue 5, pp. 1245-54, 1991 (PubMed).

Schaar, Chan, Maddox et al.: "CENP-E function at kinetochores is essential for chromosome alignment." in: The Journal of cell biology, Vol. 139, Issue 6, pp. 1373-82, 1998 (PubMed).

Validation Images
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