Phospho-Tau (T231) Rabbit Recombinant mAb
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使用信息
| 抗体应用 | WB,ELISA | ||
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| 稀释比例 |
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| 反应性 | Human Mouse Rat | ||
| MW (kDa) | 46kDa | ||
| 抗体类型 | Rabbit | ||
| 浓度 | 1mg/ml | ||
| 储存液配方 | 10 mM sodium HEPES (pH 7.5), 150 mM NaCl, 100 µg/ml BSA, 50% glycerol and less than 0.02% sodium azide. | ||
| 储存条件 (自收到货起) |
Store at –20°C. |
生物描述
| 特异性 | Phospho-Tau (T231) Rabbit Recombinant mAb detects endogenous levels of phosphorylated Tau (T231). |
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| 背景 | Tau is a neuronal protein that is critically involved in the pathogenesis of Alzheimer disease and other neurodegenerative diseases termed tauopathies. Phosphorylation of the microtubule-associated protein Tau influences the assembly and stabilization of microtubules and is deregulated in several neurodegenerative diseases. Phosphorylation of Tau generally decreases its affinity for microtubles (MTs) and abolishes its ability to stimulate MT polymerization. phosphorylation of S214 and T231 primarily decrease the ability of Tau to polymerize MTs. Phosphorylation at T231 not only regulates MT binding but is also important for the role of Tau in disease, because it detaches Tau from MTs and thus might enable the interaction with other cellular partners. Phosphorylation of T231 may also regulate the binding of SH3 domain-containing signaling proteins to the seventh PXXP motif (residues P233-P236) of Tau. Elevated phospho-tau (T231) is a marker of neurofibrillary pathology that is related to both a decrease in declarative memory and progressive atrophy of the medial temporal lobe (MTL). |
实验方法
| WB |
Western Blotting Sample preparation 1. Aspirate media from cultures and Wash the cells with 1X PBS. 2. Lyse cells by adding 1X SDS sample buffer and transfer the extract to a microcentrifuge tube. Keep onice. 3. Sonicate for 10–15 sec to complete cell lysis and shear DNA. 4. Heat a 20 µl sample to 95–100°C for 5 min, then cool on ice. 5. Centrifuge for 5 min (with Microcentrifuge). 6. Load appropriate volumes of samples onto SDS-PAGE gel (loading quantity of protein sample depends on the concentration of extracted proteins). NOTE: At the same time, please load the pre-stained molecular weight markers to determine molecular weights and verify electrotransfer. 7. Electrotransfer to nitrocellulose/PVDF membrane. Membrane Blocking and Antibody Incubations a. Blocking 1. (Optional) After transfer, wash the transferred membrane with TBS for 5 min at room temperature. 2. Incubate the membrane in the blocking buffer for 1 hr at room temperature. 3. Wash three times for 5 min each with TBST. b. Antibodies Incubation 1. Incubate membrane and primary antibody (at the appropriate dilution and diluent recommended) in a primary antibody dilution buffer with gentle agitation overnight at 4°C. 2. Wash three times for 5 min each with TBST. 3. Incubate membrane with an appropriate second antibodydissolved in the blocking buffer with gentle agitation for 1 hr at room temperature. 4. Wash three times for 5 min each with TBST. 5. Proceed with detection. Detection of Proteins 1. After antibodies incubation, Wash membrane three times for 5 minutes in TBST. 2. PrepareECL Reagent (or other chromogenic agents/substrate according to your second antibody). Mix well. 3. Incubate substrate with membrane for 1 minute, remove excess solution (membrane remains wet), wrap in plastic and expose to X-ray film. |
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