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CDKN2D 抗体

CDKN2D 适用: 人 WB, IP, IHC (p) 宿主: 小鼠 Monoclonal DCS-100 unconjugated
产品编号 ABIN487338
发货至: 中国
  • 抗原 See all CDKN2D 抗体
    CDKN2D (Cyclin-Dependent Kinase Inhibitor 2D (p19, Inhibits CDK4) (CDKN2D))
    适用
    • 49
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    宿主
    • 39
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    小鼠
    克隆类型
    • 40
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    单克隆
    标记
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    This CDKN2D antibody is un-conjugated
    应用范围
    • 39
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    Western Blotting (WB), Immunoprecipitation (IP), Immunohistochemistry (Paraffin-embedded Sections) (IHC (p))
    特异性
    This antibody reacts with Human p19INK4d
    交叉反应 (详细)
    Species reactivity (tested):Human.
    产品特性
    Synonyms: p19-INK4d, p19, INK4D, Cyclin-dependent kinase 4 inhibitor D
    纯化方法
    Protein-A Sepharose Chromatography.
    免疫原
    Bacterially produced GST-tagged fusion proteins of full-length p19INK4d. Remarks: Hybridoma was established by fusion of Mouse myeloma cell NS-2 with Balb/cmouse splenocyte.
    克隆位点
    DCS-100
    亚型
    IgG1
    Top Product
    Discover our top product CDKN2D Primary Antibody
  • 应用备注
    Western Blot: 1 μg/mLPositive Control: Jurkat cells. Immunoprecipitation: 3 μg/200-300 μL of cell extract. Positive Control: Jurkat cells. Immunohistochemistry: 1-5 μg/mLHeat treatment is necessary for Paraffin Embedded Sections. Microwave oven: 2 times for 10 minutes each in 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.
    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 the anti-p19INK4d (DCS-100) monoclonal antibody (1μg/mL) diluted with 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 Control for Western blotting: Jurkat Cells. 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-p19INK4d (DCS-100) monoclonal antibody into 250 μL of thesupernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4°C. Add 20μL of 50% Protein A-agarose beads resuspended in the Lysis buffer. Mix well and incubatewith gentle agitation 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 for10 seconds). 5) Resuspend the beads in 20 μL of Laemmli’s sample buffer, boil for 3-5 minutes, andcentrifuge for 5 minutes. Use 10 μL/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting. )Positive Controls for immunoprecipitation: Jurkat cells.
    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
    CDKN2D (Cyclin-Dependent Kinase Inhibitor 2D (p19, Inhibits CDK4) (CDKN2D))
    别名
    CDKN2D / p19INK4d (CDKN2D 产品)
    别名
    ink4d antibody, p19 antibody, p19-ink4d antibody, CDKN2D antibody, ank2 antibody, INK4D antibody, p19-INK4D antibody, INK4d antibody, p19INK4d antibody, cyclin-dependent kinase inhibitor 2D S homeolog antibody, cyclin dependent kinase inhibitor 2D antibody, cyclin-dependent kinase inhibitor 2D antibody, Cyclin-dependent kinase 4 inhibitor D antibody, cyclin-dependent kinase inhibitor 2D (p19, inhibits CDK4) antibody, similar to cyclin-dependent kinase inhibitor 2D antibody, cdkn2d.S antibody, CDKN2D antibody, cdkn2d antibody, cdn2d antibody, Cdkn2d antibody
    背景
    The INK4 family of proteins consists of four members that block progression from the G(1)-to-S phase of the cell cycle by inhibiting the activity of Cdk4 and Cdk6. p19INK4d is a 165 amino acid protein with strong structural and functional similarity to p16INK4a, a known tumor suppressor. Mutations in p19INK4d have also been associated with human osteosarcomas.Synonyms: Cyclin-dependent kinase 4 inhibitor D, INK4D, p19, p19-INK4d
    基因ID
    1032
    UniProt
    P55273
    途径
    Cell Division Cycle, Sensory Perception of Sound, Mitotic G1-G1/S Phases, Negative Regulation of intrinsic apoptotic Signaling
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