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CDC7 antibody

CDC7 Reactivity: Human WB, IHC (p), IP Host: Mouse Monoclonal DCS-342 unconjugated
Catalog No. ABIN487481
  • Target See all CDC7 Antibodies
    CDC7 (Cell Division Cycle 7 (CDC7))
    Reactivity
    • 41
    • 17
    • 12
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    Human
    Host
    • 41
    Mouse
    Clonality
    • 39
    • 2
    Monoclonal
    Conjugate
    • 19
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This CDC7 antibody is un-conjugated
    Application
    • 15
    • 15
    • 13
    • 13
    • 12
    • 6
    • 5
    • 4
    • 3
    • 2
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p)), Immunoprecipitation (IP)
    Specificity
    This antibody reacts with Human CDC7
    Cross-Reactivity (Details)
    Species reactivity (tested):Human.
    Characteristics
    Synonyms: CDC7L1, CDC7-related kinase, HsCdc7, huCdc7, Cell division cycle 7-related protein kinase
    Purification
    Protein-A Sepharose Chromatography.
    Immunogen
    Full-length Human CDC7 fusion protein. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte.
    Clone
    DCS-342
    Isotype
    IgG2b
    Top Product
    Discover our top product CDC7 Primary Antibody
  • Application Notes
    Westzern Blotting: 0.2 μg/mLImmunoprecipitation: 3 μg/200-300 μL of cell extract. Positive Controls: HeLa, Raji cells. Immunohistochemistry: 1-5 μg/mLHeat treatment is necessary for Paraffin Embedded Sections. Microwave oven: 2 times for 10 minutes each in 10 mM citrate buffer ( pH 6.5)Positive Control: Tonsil Tissue. 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 Lysisbuffer to make 8 mg/mL solution. 3) Mix the sample with equal volume of Laemmli’s sample buffer. 4) Boil the samples for 2 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 the 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 the anti-CDC7 monoclonal antibody (0. 2 μg/mL) dilutedwith 1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1: 10000 HRP-conjugated anti-mouse IgG diluted with1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 10) Wash the membrane with PBS (5 minutes x 6 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriatechemiluminescence reagents for 1 minute. Remove extra reagent from the membrane bydabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 5 minutes. Develop the film as usual. Theconditions for exposure and development may vary. Positive Controls for Western blotting: HeLa, Raji. Immunoprecipitation1) 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. 3) Add 3 μg of the anti-CDC7 monoclonal antibody into 250 μL of the supernatant. Mix welland incubate with gentle agitation for 30-120 minutes at 4°C. Add 20 μL of 50% ProteinA-agarose beads resuspended in the Lysis buffer. Mix well and incubate with gentleagitation for 60 minutes at 4°C. 4) Wash the beads 3-5 times with ice-cold Lysis buffer (centrifuge the tube at 2,500 x g for
    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 undiluted at -20 °C.
    Shelf life: one year from despatch.
    Expiry Date
    12 months
  • Target
    CDC7 (Cell Division Cycle 7 (CDC7))
    Alternative Name
    CDC7 (CDC7 Products)
    Synonyms
    CDC7L1 antibody, HsCDC7 antibody, Hsk1 antibody, huCDC7 antibody, AI597260 antibody, Cdc7l1 antibody, muCdc7 antibody, cell division cycle 7 antibody, cell division cycle 7-related protein kinase antibody, cell division cycle 7 (S. cerevisiae) antibody, CDC7 antibody, Tsp_08585 antibody, Cdc7 antibody
    Background
    Cell division cycle-7 (CDC7) was initially discovered as a temperature sensitive cell cycle mutant in Saccharomyces cerevisiae. CDC7 is required in the mitotic cell cycle for the initiation of DNA synthesis. CDC7 forms a complex with a Dbf4-related regulatory subunit to generate an active Ser/Thr kinase that functions in DNA replication, repair, and mitotic recombination. CDC7-Dbf4 efficiently phosphorylates several proteins that are required for the initiation of DNA replication, including five of the six minichromosome maintenance (MCM) proteins and the p180 subunit of DNA polymerase alpha-primase. CDC7p-Dbf4p kinase binds to chromatin during S phase and is regulated by both the APC and the RAD53 checkpoint pathway. A human homolog of the yeast CDC7 is over-expressed in some tumors and transformed cell lines.Synonyms: CDC7-related kinase, CDC7L1, Cell division cycle 7-related protein kinase, HsCdc7, huCdc7
    Gene ID
    8317
    UniProt
    O00311
    Pathways
    Mitotic G1-G1/S Phases, DNA Replication
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