| 名稱 | DBEQ |
| 描述 | DBEQ (JRF 12) is a selective, potent, reversible, and ATP-competitive p97 inhibitor. |
| 細(xì)胞實(shí)驗(yàn) | Cells are seeded on a 384-well solid white plate (5,000 cells/well). Cells are transfected with luciferase siRNA or p97 siRNA (10 nM) for 48 hours or treated with DBeQ for the indicated amount of time. Caspase-3/7 Glo, caspase-6 Glo, caspase-8 Glo, or caspase-9 Glo is added into each well and mixed by shaking at 500 rpm for 1 min. Luminescence signal is determined after incubation at room temperature for 1 hour. Cellular viability is determined with CellTiter-Glo reagen. To determine the IC50 of cellular viability, cells are treated with MG132 or DBeQ at seven concentrations (threefold serial dilutions starting at 33 μM) for 48 hours. IC50 values are calculated from ?tting the percentage of luminescence signal normalized to DMSO treated cells).(Only for Reference) |
| 激酶實(shí)驗(yàn) | Manual ATPase Assay: Assay Buffer [20 μL of 2.5× concentration, where 1× = 50 mM Tris (pH 7.4), 20 mM MgCl2, 1 mM EDTA, and 0.5 mM tris(2-carboxyethyl)phosphine (TCEP)] is dispensed into each well of a 96-well plate. Puri?ed p97 (25 μL of 50 μM) is diluted in 975 μL of 1× Assay Buffer, and 10 μL is dispensed in each well. DBeQ (10 μL) or 5% DMSO (10 μL) is then added to each well, and the plate is incubated at room temperature for 10 min. The ATPase assay is carried out by adding to each well 10 μL of 500 μM ATP (pH 7.5), incubating at room temperature for 60 min, and then adding 50 μL Kinase Glo Plus reagent, followed by a ?nal 10-min incubation at room temperature in the dark. Luminescence is read on an Analyst AD. DBeQ is assayed at a range of concentrations (0, 0.048, 0.24, 1.2, 6, and 30 μM) in triplicate. |
| 體外活性 | 10 μM DBeQ完全抑制病毒和抗體的降解,但不抑制IgG Fc的降解。10 μM DBeQ快速促進(jìn)HeLa細(xì)胞中caspases-3和-7激活。比激活外在caspase-8通路,DBeQ更有效激活內(nèi)在caspase-9凋亡途徑,而STS激活兩種途徑程度相似。10 μM DBeQ有效抑制HEK293細(xì)胞中TCRα-GFP降解。DBeQ作用于HEK293細(xì)胞,3小時(shí)內(nèi)劑量依賴性誘導(dǎo)CHOP,但不增加p21水平。DBeQ作用于U20S細(xì)胞,降低基本的和營(yíng)養(yǎng)刺激的MTOR靶點(diǎn)磷酸化,與雷帕霉素效果類似。15 μM DBeQ 作用于Hela細(xì)胞,誘導(dǎo)LC3-II 在在細(xì)胞核及濃縮膜和胞質(zhì)級(jí)組分中大量積累。DBeQ作用于HeLa細(xì)胞,抑制UbG76V-GFP,ODD-Luc和Luc-ODC 降解,IC50分別為2.6 μM、56 μM 和45 μM。 |
| 存儲(chǔ)條件 | Powder: -20°C for 3 years | In solvent: -80°C for 1 year | Shipping with blue ice. |
| 溶解度 | DMSO : 3.85 mg/mL (11.3 mM), Sonication is recommended.
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| 關(guān)鍵字 | Inhibitor | DBEQ | Autophagy | p97 | VCP | JRF12 | inhibit | JRF-12 | Apoptosis | Cdc48 |
| 相關(guān)產(chǎn)品 | Guanidine hydrochloride | Naringin | Valproic Acid | L-Glutamic acid | Gefitinib | Hydroxychloroquine | Dextran sulfate sodium salt (MW 4500-5500) | Stavudine | Tributyrin | L-Ascorbic acid | Paeonol | Sodium 4-phenylbutyrate |
| 相關(guān)庫(kù) | 抑制劑庫(kù) | 經(jīng)典已知活性庫(kù) | 已知活性化合物庫(kù) | 自噬庫(kù) | 抗衰老化合物庫(kù) | HIF-1化合物庫(kù) | NO PAINS 化合物庫(kù) | 抗病毒庫(kù) | 泛素化化合物庫(kù) | 內(nèi)質(zhì)網(wǎng)應(yīng)激化合物庫(kù) |