CRADD 抗体
Quick Overview for CRADD 抗体 (ABIN5540081)
抗原
See all CRADD 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 RAIDD.
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交叉反应 （详细）
- Does not react with Mouse (WR19L, Ba/F3), Rat (PC12, Rat-1), and Hamster (BHK).
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纯化方法
- Protein A agarose
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免疫原
- Recombinant full-length human RAIDD (200 a.a.)
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亚型
- IgG1
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anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 1-199) antibody
CRADD 适用: 人 WB, ELISA, IF 宿主: 小鼠 Monoclonal 1F8 unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 1-199) antibodyCRADD 适用: 人 ELISA, IHC 宿主: 兔 Polyclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 1-199) antibodyCRADD 适用: 人 WB, ELISA, ICC, IHC (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 1-199) antibodyCRADD 适用: 人 WB 宿主: 兔 Polyclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) antibodyCRADD 适用: 人 WB, IHC 宿主: 兔 Monoclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) antibodyCRADD 适用: 人 WB, IF 宿主: 兔 Monoclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 99-117), (Middle Region) antibodyCRADD 适用: 人 WB, IF, EIA 宿主: 兔 Polyclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 1-199) antibodyCRADD 适用: 人 ELISA, IF 宿主: 兔 Polyclonal unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) (AA 1-199) antibodyCRADD 适用: 人 WB, IHC, ICC, IP 宿主: 小鼠 Monoclonal 23# unconjugated
anti-CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD) antibody (HRP)CRADD 适用: 人 WB, ELISA, IHC, IHC (p) 宿主: 兔 Polyclonal HRP
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应用备注
- Western blot: 1 μg/mL for chemiluminescence detection system. 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 volumes 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 cold Lysis buffer to make an 8 mg/mL solution. 3) Mix the sample with an equal volume of Laemmli's sample buffer. 4) Boil the samples for 3 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 specific 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 suggested in the APPLICATIONS for 1 hour at room temperature. (The optimal antibody concentration will depend on the experimental conditions.) 8) Wash the membrane with PBS-T [0.05 % Tween-20 in PBS] (10 minutes x 3 times). 9) Incubate the membrane with the 1:10,000 HRP-conjugated anti-mouse IgG diluted with 1 % skimmed milk (in PBS, pH 7.2) for 1 hour at room temperature. 10) Wash the membrane with PBS-T (10 minutes x 3 times). 11) Wipe excess buffer from the membrane, then incubate it with appropriate chemilum inescence reagents for 1 minute. Remove extra reagent from the membrane by dabbing with a paper towel, and seal it in plastic wrap. 12) Expose to an X-ray film in a dark room for 1 minute. Develop the film as usual. The conditions for exposure and development may vary. (Positive controls for Western blotting Jurkat, HeLa)
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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. Contains no preservatives.
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储存液
- Without preservative
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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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- CRADD (CASP2 and RIPK1 Domain Containing Adaptor with Death Domain (CRADD))
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别名
- raidd
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背景
- Apoptosis is a major form of cell death characterized by severa l morphological features that include chromatin condensation and fragmentation, cell membrane blebbing, and formation of apoptotic bodies. These morphological changes occur via signaling pathway that leads to the recruitment and activation of caspases, a family of cysteine-containing , aspartate-specific proteases. Adaptor molecules that contai n protein-protein interaction motifs mediate the coupling of caspases to signaling pathway. For example, in the TNF receptor I mediated apoptosis pathway, the apoptotic signal is transduced through interaction of the oligomerized receptor death domain (DD) with a set of cy toplasmic adaptor/signaling molecules including FADD, TRADD and RIP. RAIDD (also known as CRADD) is another candidate adaptor molecule. It has a bipartite architecture comprising a carboxy-terminal DD and an amino-terminal caspase recruitment domain (CARD) highly homologous with the sequence of the prodomain of human caspase-2 and C. elegans CED-3. RAIDD interacts with RIP on respective DD, and with caspase-2 or CED-3 on respective CARD. Thus it may play a role in the TNF-receptor I mediated apoptosis by recruiting caspase-2 to the pathway. However, it has been remained unclea r about the significance of caspase-2 activation through the RIP/RAIDD pathway for death induction at least in some situation, because targeted disruption of RIP in mice, as well as mutational ablation of RIP function in cultured cells did not interfere with death induction by TNF nor by Fas, while NF- κ B activation through RIP was abolished in these cells.
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UniProt
- P78560
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途径
- Apoptosis, Caspase Cascade in Apoptosis, Positive Regulation of Endopeptidase Activity
抗原
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