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p53 抗体 (pSer315)

TP53 适用: 人 WB, IP 宿主: 小鼠 Monoclonal FPS315 unconjugated
产品编号 ABIN487474
发货至: 中国
  • 抗原 See all p53 (TP53) 抗体
    p53 (TP53) (Tumor Protein P53 (TP53))
    抗原表位
    • 43
    • 39
    • 37
    • 30
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    • 3
    pSer315
    适用
    • 823
    • 252
    • 208
    • 104
    • 47
    • 33
    • 31
    • 18
    • 10
    • 9
    • 7
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    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    宿主
    • 623
    • 217
    • 6
    • 2
    • 1
    小鼠
    克隆类型
    • 559
    • 289
    • 1
    单克隆
    标记
    • 476
    • 38
    • 31
    • 19
    • 18
    • 13
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    • 2
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    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    This p53 antibody is un-conjugated
    应用范围
    • 661
    • 298
    • 247
    • 200
    • 199
    • 148
    • 124
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    • 119
    • 64
    • 59
    • 32
    • 18
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    • 9
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    • 3
    • 3
    • 2
    • 2
    • 2
    • 1
    • 1
    • 1
    • 1
    • 1
    • 1
    Western Blotting (WB), Immunoprecipitation (IP)
    特异性
    This antibody reacts with Human p53 phosphorylated at Ser315.
    交叉反应 (详细)
    Species reactivity (tested):Human.
    产品特性
    Synonyms: Cellular tumor antigen p53, Tumor suppressor p53, Phosphoprotein p53, NY-CO-13
    纯化方法
    Protein-A Sepharose Chromatography
    免疫原
    A 15-amino acid phosphopeptideAA Sequence: RALPNNTSSpSPQPKKKbRemarks: Hybridoma was established by fusion of mouse myeloma cell SP-2 with Balb/cmouse splenocytes.
    克隆位点
    FPS315
    亚型
    IgG1
    Top Product
    Discover our top product TP53 Primary Antibody
  • 应用备注
    Immunoprecipitation/Western blotting: 5 μg/mLPositive Control: Raji cells. Detailed procedure is provided in Protocols. This monoclonal antibody should be used by Western blot in combination withImmunoprecipitation with anti-p53 (DO-1). This antibody is not recommended for use by Western blotting using crude cell lysates.
    Other applications not tested.
    Optimal dilutions are dependent on conditions and should be determined by the user.
    实验流程
    Immunoprecipitation / SDS PAGE & Western Blotting1) Wash the cells 3 times with PBS and suspend with 10 volumes of cold Lysis buffer (50mM Tris-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 100 µL of the anti-p53 monoclonal antibody into 200 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 for10 seconds). 5) Resuspend the beads in 20 ìL of Laemmli’s sample buffer, boil for 3-5 minutes, andcentrifuge for 5 minutes. Load 10 µL of the sample per lane in a 1 mm thickSDS-polyacrylamide gel for electrophoresis. 6) 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 specific transfer procedure. 7) 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. 8) Incubate the membrane with the anti-phospho p53 Ser315 monoclonal antibody (5µg/mL) diluted with 1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 9) Wash the membrane with PBS (5 minutes x 6 times). 10) Incubate the membrane with the 1: 10000 POD-conjugated anti-mouse IgG ê dilutedwith 1% skimmed milk (in PBS, pH 7. 2) for 1 hour at room temperature. 11) Wash the membrane with PBS (5 minutes x 6 times)12) 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. 13) Expose to 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 Immunoprecipitation/Western blotting: Raji 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
    4 °C
    储存方法
    Store the antibody undiluted at 2-8 °C.
    Shelf life: one year from despatch.
    有效期
    12 months
  • 抗原
    p53 (TP53) (Tumor Protein P53 (TP53))
    别名
    p53 (TP53) (TP53 产品)
    背景
    P53 tumor suppressor proteins play a critical role in cellular defense against DNA-damaging reagents. Activation of p53 causes cell cycle arrest and sometimes, apoptosis, resulting in the prevention of genetically damaged cells from proliferating. DNA damage increases p53 protein levels and activates transcription of the p21 gene. The p21 protein binds to and inhibits the cell cycle regulator cdc2 kinase, causing G1 arrest. p53 is regulated by phosphorylation at multiple sites by several different protein kinases. The cdc2 kinase phosphorylates Ser315 of human p53 in vitro, and the majority of p53 protein is phosphorylated at Ser315 following irradiation damage.Synonyms: Cellular tumor antigen p53, NY-CO-13, Phosphoprotein p53, Tumor suppressor p53
    基因ID
    7157
    UniProt
    P04637
    途径
    p53 Pathway, MAPK Pathway, PI3K-Akt Signaling, Apoptosis, AMPK Signaling, Chromatin Binding, ER-Nucleus Signaling, Positive Regulation of Endopeptidase Activity, Hepatitis C, Protein targeting to Nucleus, Autophagy, Warburg Effect
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