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CDC27 antibody (C-Term)

CDC27 Reactivity: Human, Rat, Mouse, Dog, Cow, Xenopus laevis WB, IP, IHC (p), EIA Host: Mouse Monoclonal AF3-1 unconjugated
Catalog No. ABIN492595
  • Target See all CDC27 Antibodies
    CDC27 (Cell Division Cycle 27 Homolog (S. Cerevisiae) (CDC27))
    Binding Specificity
    • 16
    • 15
    • 6
    • 5
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    AA 814-823, C-Term
    Reactivity
    • 67
    • 43
    • 25
    • 7
    • 6
    • 6
    • 5
    • 4
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Human, Rat, Mouse, Dog, Cow, Xenopus laevis
    Host
    • 79
    • 4
    Mouse
    Clonality
    • 72
    • 11
    Monoclonal
    Conjugate
    • 31
    • 5
    • 4
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 3
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    This CDC27 antibody is un-conjugated
    Application
    • 27
    • 27
    • 25
    • 23
    • 10
    • 10
    • 7
    • 7
    • 4
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Enzyme Immunoassay (EIA)
    Specificity
    This antibody reacts with 97 kDa membrane protein CDC27 on Western blotting, Immunoprecipitation and Immunohistochemistry.
    Cross-Reactivity (Details)
    Species reactivity (expected):Bovine, Canine and Xenopus leavis.
    Species reactivity (tested):Human, Mouse, Rat.
    Characteristics
    Synonyms: CDC27Hs, H-NUC, D0S1430E, D17S978E, Cell division cycle protein 27 homolog
    Purification
    Protein-A Agarose Chromatography of hybridoma supernatant.
    Immunogen
    Peptide corresponding to C-terminal of Human CDC27 (aa 814-823).
    Clone
    AF3-1
    Isotype
    IgG2b
    Top Product
    Discover our top product CDC27 Primary Antibody
  • Application Notes
    Western blotting: 1 μg/mL for chemiluminescence detection system. Positive Controls: Jurkat, Raji, K562, HL-60. Immunoprecipitation: 2 μg/200 μL of cell extract. Positive Control: Raji. Immunohistochemistry on Paraffin Embedded Sections: 10 μg/mL. (Heat treatment is necessary)Autoclave: 10 minutes at 110 °C in 10 mM citrate buffer ( pH 6.5). Positive Control: Tonsil. Detailed procedure is provided in Protocols.
    Other applications not tested.
    Optimal dilutions are dependent on conditions and should be determined by the user.
    Protocol
    SDS-PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl, pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10%glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. Measure the protein concentration of the supernatant and add the cold Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 minutes and centrifuge. Load 10 μL of the sample per lane in a 1mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm2 for 1 hourin a semi-dry transfer system (Transfer Buffer: 25 mM Tris, 190 mM glycine, 20% MeOH). See the manufacture's manual for precise transfer procedure. 6) To reduce nonspecific binding, soak the membrane in 10% skimmed milk (in PBS, pH7. 2) for 1 hour at room temperature, or overnight at 4°C. 7) Incubate the membrane with primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggest in the APPLICATIONS for 1 hour at room temperature. (Theconcentration of antibody will depend on condition. )8) Wash the membrane with PBS-T [0. 05% Tween-20 in PBS] (5 minutes x 3 times). 9) Incubate the membrane with the 1: 10,000 HRP-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at RT. 10) Wash the membrane with PBS-T (10 minutes x 3 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriatechemiluminescence reagent for 1 minute. 12) Remove extra reagent from the membrane by dabbing with paper towel, and seal it inplastic wrap. 13) Expose to an X-ray film in a dark room for 3 minutes. 14) Develop the film as usual. The condition for exposure and development may vary. Positive Controls: Jurkat, Raji, K562, HL-60, MCF-7Immunoprecipitation1) Wash the cells 3 x with PBS and suspend with 10 volume of cold Lysis buffer (50 mMTris-HCl pH 7. 2, 250 mM NaCl, 0. 1% NP-40, 2 mM EDTA, 10% glycerol) containingappropriate protease inhibitors. Incubate it at 4°C with rotating for 30 minutes, thensonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4°C and transfer the supernatant toanother tube. 3) Add primary antibody as suggested in the APPLICATIONS into 200 µL of the supernatant.
    Restrictions
    For Research Use only
  • Concentration
    1.0 mg/mL
    Buffer
    PBS, pH 7.2 containing 50 % Glycerol without preservatives.
    Preservative
    Without preservative
    Storage
    -20 °C
    Storage Comment
    Store the antibody (in aliquots) at -20 °C. Avoid repeated freezing and thawing.
    Shelf life: one year from despatch.
    Expiry Date
    12 months
  • Target
    CDC27 (Cell Division Cycle 27 Homolog (S. Cerevisiae) (CDC27))
    Alternative Name
    CDC27 (CDC27 Products)
    Background
    CDC16, CDC27 and CDC34 are components of the APC (anaphase-promoting complex) that also play a role as components of the ubiquitin- conjugating E3 enzyme. A 20S complex containing CDC27 and CDC16 catalyzes the mitosis-specific conjugation of ubiquitin to cyclin B, resulting in cyclin B/Cdk complex degradation. CDC27 is therefore required for progression of the cell cycle, where it executes essential mitotic functions near the metaphase/anaphase transition. CDC27 is ~97 kDa and is localized to the centrosome and mitotic spindle.Synonyms: CDC27Hs, Cell division cycle protein 27 homolog, D0S1430E, D17S978E, H-NUC
    Gene ID
    996
    UniProt
    P30260
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