TRAF6 抗体
Quick Overview for TRAF6 抗体 (ABIN5541395)
抗原
See all TRAF6 抗体适用
宿主
克隆类型
标记
应用范围
克隆位点
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特异性
- This antibody reacts with TRAF6 (60 kDa).
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纯化方法
- Protein G agarose
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免疫原
- Recombinant mouse TRAF6
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亚型
- IgG
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anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 153-423) antibody
TRAF6 适用: 人 WB, IHC, IF, ICC 宿主: 兔 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 413-522) antibodyTRAF6 适用: 人 WB, ELISA, RNAi, PLA 宿主: 小鼠 Monoclonal 1B2 unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 1-522) antibodyTRAF6 适用: 人 IHC, ELISA, IF 宿主: 兔 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 263-522) antibodyTRAF6 适用: 人 WB, IF 宿主: 兔 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (Middle Region) antibodyTRAF6 适用: 人, 小鼠, 大鼠, Cow, Pig, 兔, 马 WB 宿主: 兔 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 441-530) antibodyTRAF6 适用: 人, 小鼠, 大鼠, 小鸡 WB, ELISA, IF (cc), IF (p), IHC (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 314-343) antibodyTRAF6 适用: 人 WB 宿主: 兔 Polyclonal RB40708 unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 1-522) antibodyTRAF6 适用: 人 WB, IP 宿主: 兔 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (AA 1-522) antibodyTRAF6 适用: 人 WB, IF 宿主: 小鼠 Polyclonal unconjugated
anti-TNF Receptor-Associated Factor 6 (TRAF6) (C-Term) antibodyTRAF6 适用: 人, 小鼠 WB, IHC, IP, IHC (p) 宿主: 兔 Polyclonal unconjugated
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应用备注
- Western blot: 1 μg/200 mL for chemiluminescence detection system. Immunoprecipitation: 1 μg/200 μL of cell extract from 5x10 6 cells. For details see protocol below.
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实验流程
- SDS-PAGE & Western blotting 1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris-HCl, pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. Measure the protein concentration of the supernatant and add the Lysis buffer 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 μ L of the sample per lane in a 1 mm thick SDS-polyacrylamide gel for electrophoresis. 5) Blot the protein to a polyvinylidene difluoride (PVDF) membrane at 1 mA/cm 2 for 1 hour in 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, pH 7.2) for 1 hour at room temperature, or overnight at 4 o 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. (The concentration of antibody will depend on condition.) 8) Wash the membrane with PBS-T [0.05 % Tween-20 in PBS] (5 minutes x 6 times). 9) Incubate the membrane with the 1:1,000 HRP-conjugated anti-Hamster IgG diluted with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS-T (5 minutes x 6 times). 11) Wipe excess buffer on the membrane, then incubate it with appropriate chemiluminescence reagent for 1 minute. Remove extra reagent from the membrane by dabbing with 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. The condition for exposure and development may vary. (Positive controls for Western blotting L5178Y, WR19L, NIH/3T3) Immunoprecipitation 1) Wash the cells 3 times with PBS and suspend with 10 volume of cold Lysis buffer (50 mM Tris-HCl pH 7.2, 250 mM NaCl, 0.1 % NP-40, 2 mM EDTA, 10 % glycerol) containing appropriate protease inhibitors. Incubate it at 4 o C with rotating for 30 minutes, then sonicate briefly (up to 10 seconds). 2) Centrifuge the tube at 12,000 x g for 10 minutes at 4 o C and transfer the supernatant to another tube. 3) Add primary antibody as suggest in the APPLICATIONS into 200 μ L of the supernatant. Mix well and incubate with gentle agitation for 30-120 minutes at 4 o C. Add 20 μ L of 50 % protein G agarose beads resuspended in the cold Lysis buffer. Mix well and incubate with gentle agitation for 60 minutes at 4 o C. 4) Wash the beads 3-5 times w ith the cold Lysis buffer (centrifuge the tube at 2,500 x g for 10 seconds). 5) Resuspend the beads in 20 μL of Laemmli's sample buffer, boil for 3-5 minutes, and centrifuge for 5 minutes. Use 10 μL/lane for the SDS-PAGE analysis. (See SDS-PAGE & Western blotting.) (Positive control for Immunoprecipitation L5178Y)
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限制
- 仅限研究用
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状态
- Liquid
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缓冲液
- PBS containing 50 % glycerol, pH 7.2. No preservative is contained.
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储存液
- Azide free
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储存条件
- -20 °C
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储存方法
- Upon receipt, store undiluted (in aliquots) at -20°C. Avoid repeated freezing and thawing. Shelf life: One year from despatch.
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- TRAF6 (TNF Receptor-Associated Factor 6 (TRAF6))
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别名
- traf6,rnf85
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背景
- TRAFs (TNF receptor associated factors) form a family of cyt oplasmic adaptor proteins that mediate signal transduction from many members of the TNF-receptor superfamily. TR AF6 also participates in signal transduction of the IL-1R/Toll receptor superfamily. TRAF6 is a ~55 kDa cytoplasmi c adapter protein that plays diverse roles in perinatal and postnatal survival, bone metabolism, adaptive immunity, cytokine signaling, and the regulation of inflammatory and apoptotic responses through activation of the transcription factors of NF- κ B. TRAF6 overexpression activates MAP kinases, including ERK, p38 and JNK, and TRANCE/OPGL Akt/PKB through a signaling complex involving c-Src and TRAF6. TRAF6 dimerizes strongly with itself and weakly with TRAF2 and TRAF3.
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UniProt
- P70196
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途径
- NF-kappaB Signaling, TCR Signaling, TLR signaling, Fc-epsilon Receptor Signaling Pathway, Neurotrophin Signaling Pathway, Activation of Innate immune Response, Regulation of Leukocyte Mediated Immunity, Positive Regulation of Immune Effector Process, Production of Molecular Mediator of Immune Response, Tube Formation, Hepatitis C, Toll-Like Receptors Cascades, Ubiquitin Proteasome Pathway
抗原
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