Caspase 3 抗体
Quick Overview for Caspase 3 抗体 (ABIN5541795)
抗原
See all Caspase 3 (CASP3) 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 Pro-Caspase-3 (32 kDa) and active Caspase-3 (17 kDa).
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纯化方法
- Protein G agarose
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免疫原
- Full length human recombinant Caspase-3
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亚型
- IgG2a
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anti-Caspase 3 (CASP3) (AA 60-100) antibody
CASP3 适用: 人, 小鼠, 大鼠, Cow, 山羊 WB, IF (cc) 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (AA 67-175) antibodyCASP3 适用: 人, 小鼠, 大鼠 WB, IHC, ICC, IF 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (AA 220-236), (C-Term) antibodyCASP3 适用: 人 WB, IHC, ICC 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (AA 11-120) antibodyCASP3 适用: 人, 小鼠, 大鼠 WB, ELISA, IHC (p), FACS, IF (cc), IF (p), IHC (fro) 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) antibodyCASP3 适用: 人, 小鼠, 大鼠 WB, IHC 宿主: 小鼠 Monoclonal 5E1 unconjugated
anti-Caspase 3 (CASP3) (cleaved) antibodyCASP3 适用: 人, 小鼠, 大鼠 IHC, IF 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (Internal Region) antibodyCASP3 适用: 人, 小鼠, 大鼠, Cow WB, IHC, ELISA, ICC, IF 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (pSer150) antibodyCASP3 适用: 人, 小鼠, 大鼠 WB, IHC, ELISA, ICC, IF 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (AA 29-175) antibodyCASP3 适用: 人 WB, IHC 宿主: 兔 Polyclonal unconjugated
anti-Caspase 3 (CASP3) (AA 29-175) antibodyCASP3 适用: 人 WB, IHC, IP, ICC 宿主: 小鼠 Monoclonal C3 unconjugated
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应用备注
- Western blot: 1 μg / ml for chemiluminescence detection system. For details see protocol below. Not recommended for Immunoprecipitatoin or Immunohistochemistry.
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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 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 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 (5 minutes x 6 times). 9) Incubate the membrane with the 1:10,000 HRP-conjugated anti-rat IgG diluted with 1 % 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 appropriate chemiluminescence 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 X-ray film in a dark room for 5 minutes. Develop the film as usual. The conditions for exposure and development may vary. Positive controls for Western blotting Jurkat and Apoptotic Jurkat
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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.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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- Caspase 3 (CASP3)
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别名
- caspase-3
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背景
- Caspase-3 (also known as CPP32, Yama, and apopain) is a key member of the caspase family of cysteine proteases. Caspase-3 exists in cells as an inactive 32 kDa proenzyme. During apoptosis pro-caspase-3 is processed at aspartate residues by self-proteolysis and/or cleav age by upstream caspases, such as caspase6, 8, or 9. The processed form of caspase-3 consists of large (17 kDa) and small (12 kDa) subunits which associate to form the active tetrameric enzyme tetramer (a pair of heterodimers). The active caspase-3 proteolytically cl eaves and activates other caspases, as well as relevant targets in the cells (e.g., PARP, SREBPs, and DFF). Activation of pro-caspase-3 stands at a point of convergence for the two major types of apoptosis signaling pathways-those linked to cell surface death receptors and th ose linked to mitochondrial release of cytochrome c.
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UniProt
- P42574
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途径
- Apoptosis, Caspase Cascade in Apoptosis, Sensory Perception of Sound, ER-Nucleus Signaling, Positive Regulation of Endopeptidase Activity, Activated T Cell Proliferation
抗原
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