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CDC25C 抗体

This anti-CDC25C antibody is a 小鼠 单克隆 antibody detecting CDC25C in WB. Suitable for 人, 小鼠 和 大鼠.
产品编号 ABIN487484
发货至: 中国

Quick Overview for CDC25C 抗体 (ABIN487484)

抗原

See all CDC25C 抗体
CDC25C (Cell Division Cycle 25 Homolog C (S. Pombe) (CDC25C))

适用

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人, 小鼠, 大鼠

宿主

  • 102
  • 15
小鼠

克隆类型

  • 98
  • 19
单克隆

标记

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This CDC25C antibody is un-conjugated

应用范围

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Western Blotting (WB)

克隆位点

DCS-193
  • 特异性

    This antibody reacts with CDC25C (60 kDa) on Western blotting.

    交叉反应 (详细)

    Species reactivity (tested):Human, Mouse and Rat.

    产品特性

    Synonyms: M-phase inducer phosphatase 3, Dual specificity phosphatase Cdc25C, Cell Division Cycle25C

    纯化方法

    Protein-A Agarose Chromatography

    免疫原

    Human CDC25C. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte.

    亚型

    IgG1
  • 应用备注

    Western Blot: 10 μg/mL for chemiluminescence detection system. Positive Controls: HeLa, A431, ZR75-1, HEp-2, NIH/3T3, PC12. Detailed procedure is provided in Protocols.
    Other applications not tested.
    Optimal dilutions are dependent on conditions and should be determined by the user.

    实验流程

    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 primary antibody diluted with PBS, pH 7. 2 containing 1%skimmed milk as suggested in the APPLICATIONS for 1 hour at room temperature. (Theoptimal antibody concentration will depend on the experimental conditions. 8) Wash the membrane with PBS (5 minutes x 6 times). 9) Incubate the membrane with the 1: 10,000 POD-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, A431, ZR75-1, HEp-2, NIH/3T3, PC12

    限制

    仅限研究用
  • 浓度

    1.0 mg/mL

    缓冲液

    PBS, pH 7.2 containing 50 % Glycerol without preservatives

    储存液

    Without preservative

    储存条件

    -20 °C

    储存方法

    Store the antibody undiluted at -20 °C.
    Shelf life: one year from despatch.

    有效期

    12 months
  • 抗原

    CDC25C (Cell Division Cycle 25 Homolog C (S. Pombe) (CDC25C))

    别名

    CDC25C

    背景

    The activity of cyclin-dependent kinases is regulated by their phosphorylation status, which is controlled by the antagonistic action of CDC25 phosphatases. Three CDC25 genes are present in human cells: CDC25A, CDC25B and CDC25C. CDC25C is a ~56 kDa dual specific protein phosphatase that controls entry into mitosis by regulating the dephosphorylation of the Cdk1/cyclin B complex. Phosphorylation of Cdc25C creates a binding site for 14-3-3 protein which inhibits Cdc25C. This prevents activation of the Cdk1/cyclin B complex and prevents mitotic entry. Cdc25C is phosphorylated by checkpoint kinases throughout interphase but becomes dephosphorylated during mitosis. Cdc25C is also regulated by p53 via a p53 response element in its promoter, and it is predominantly expressed in the G2 phase of the cell cycle.Synonyms: Cell Division Cycle 25C, Dual specificity phosphatase Cdc25C, M-phase inducer phosphatase 3

    基因ID

    995

    UniProt

    P30307

    途径

    Cell Division Cycle, M Phase
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