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DAXX 抗体 (C-Term)

This anti-DAXX antibody is a 小鼠 单克隆 antibody detecting DAXX in WB 和 IF. Suitable for 人.
产品编号 ABIN501203
发货至: 中国

Quick Overview for DAXX 抗体 (C-Term) (ABIN501203)

抗原

See all DAXX 抗体
DAXX (Death-Domain Associated Protein (DAXX))

适用

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宿主

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小鼠

克隆类型

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单克隆

标记

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This DAXX antibody is un-conjugated

应用范围

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Western Blotting (WB), Immunofluorescence (IF)

克隆位点

DAXX-01
  • 抗原表位

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    AA 558-740, C-Term

    特异性

    This antibody reacts with Daxx (110 kDa).

    交叉反应 (详细)

    Species reactivity (tested):Human.

    纯化方法

    Protein-A Sepharose Chromatography of hybridoma supernatant.

    免疫原

    Recombinant C-terminal part (aa 558-740) of Human Daxx

    亚型

    IgG1
  • 应用备注

    Western blotting: 1 μg/mL for chemiluminescence detection system. Immunocytochemistry: 10 μg/mL. 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 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. Immunocytochemistry1) Culture the cells in the appropriate condition on a glass slide. (for example, spread 10e4of HEp-II cells for one slide, then incubate in a CO2 incubator for one night. )2) Fix the cells by immersing the slide in MeOH containing methanol for 10 minutes on ice. 3) Air dry the slides. 4) Add the primary antibody diluted with PBS as suggest in the APPLICATIONS onto thecells and incubate for 1 hour at room temperature (Optimization of antibody concentrationor incubation condition are recommended if necessary). 5) Prepare a wash container such as a 500 mL beaker with a stirrer. Then wash the culturedcells on the glass slide by soaking the slide with a plenty of PBS in the wash container for 5minutes. Take care not to touch the cells. Repeat another washes once more. 6) Add the secondary antibody 1: 100 FITC conjugated anti-mouse IgG onto the cells. Incubate for 1 hour at room temperature. Keep out light by aluminum foil. 7) Wash the slide in a plenty of PBS as in the step 5). 8) Wipe excess liquid from slide but take care not to touch the cells. Never leave the cellsto dry. 9) Promptly add PermafluorTM aqueous mounting medium onto the slide, then put a cover

    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

    注意事项

    Avoid repeated freezing and thawing.

    储存条件

    -20 °C/-80 °C

    储存方法

    Store lyophilized (preferably in a desiccator) at -20 °C and in aliquots at -80 °C. Reconstituted antibody can be stored at 4 °C for a limited period of time, it does not show decline in activity after two weeks at 4 °C.
  • 抗原

    DAXX (Death-Domain Associated Protein (DAXX))

    别名

    DAXX

    背景

    Daxx binds specifically to the cytosolic Death Domain (DD) of the Fas receptor and enhances Fas-induced apoptosis. Daxx also links this receptor to an apoptosis pathway involving activation of c-Jun N-terminal kinase (JNK). Daxx is widely expressed in fetal and adult tissues, indicating its important function in Fas signaling pathways. The Daxx protein resides primarily in the nucleus but it is sequestered in the cytoplasm by ASK1 (apoptosis signal-regulating kinase 1), resulting in inhibition of Daxx-mediated transcriptional regulation.Synonyms: BING2, DAP6, Death domain-associated protein 6, EAP1, ETS1-associated protein 1, Fas death domain-associated protein

    基因ID

    1616

    NCBI登录号

    NP_001135441

    UniProt

    Q9UER7

    途径

    Intracellular Steroid Hormone Receptor Signaling Pathway
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