用户验证的应用
经验证应用
| 项目 | 详情 |
|---|---|
| WB | Western blot analysis of extracts from HeLa (lane 1(20μg)) 、HepG2 (lane 2(20μg)) 、A549 (lane 3(20μg)) 、SH-SY5Y (lane 4(20μg)) 、HEK293 (lane 5(20μg)) 、Jurkat (lane 6(20μg)) and Mouse testis (lane 7(20μg)) using SirT1 Antibody. Proteins were transferred to a PVDF membrane and blocked with 5% nonfat dry milk in TBST for 1.5 hour at room temperature. The primary antibody (HY-P80319, 1/1000), competitor's antibody (1/1000) and Loading control antibody (GAPDH, 1/10000) was used in 5% nonfat dry milk in TBST at 4℃ overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature. Western blot analysis of extracts from HEK293(lane 2(20ug), A549(lane 3(20ug) and Hela(lane 4(20ug) using SIRT1 Antibody ( HY-P80319 ) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P83730, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse/Rabbit IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature. Western blot analysis of extracts from HeLa (lane 1(20μg)) 、HepG2 (lane 2(20μg)) 、A549 (lane 3(20μg)) 、SH-SY5Y (lane 4(20μg)) 、HEK293 (lane 5(20μg)) 、Jurkat (lane 6(20μg)) and Mouse testis (lane 7(20μg)) using SirT1 Antibody ( HY-P80319 ). Proteins were transferred to a PVDF membrane and blocked with 5% nonfat dry milk in TBST for 1.5 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (GAPDH, 1/10000) was used in 5% nonfat dry milk in TBST at 4℃ overnight. Goat Anti-Rabbit IgG-HRP Secondary Antibody (1/10,000) was used for 1 hour at room temperature. Western blot analysis was performed on protein extracts (30 μg) from HT-1080 (lane 2), SH-SY5Y (lane 3), HeLa (lane 4), HepG2 (lane 5), A549 (lane 6), and Caco-2 (lane 7) using SIRT1 antibody. Proteins were transferred onto a 0.45 μm PVDF membrane using the Trans-Blot ® Turbo ™ system for 13 min. The membrane was then blocked with 5% nonfat milk in TBST ( HY-K1025 ) for 1 h at room temperature. Thhe primary antibody (1:1000) and loading control antibody GAPDH Antibody (HRP) ( HY-P80954A ) (1:2500) were diluted in 5% nonfat milk in TBST and incubated with the membrane overnight at 4°C. After washing, the membrane of primary antibody was incubated with HRP-conjugated goat anti-rabbit IgG (H&L) secondary antibody ( HY-P8001 ) (1:5000) diluted in 5% nonfat milk in TBST for 1 h at room temperature. Protein bands were visualized using an Ultra High Sensitivity ECL detection kit ( HY-K1005 ). |
| IHC-P | Immunohistochemical analysis of paraffin-embedded rat testis tissue using SIRT 1 Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P80319, 1/200), competitor's antibody (1/200) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam. Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue using SIRT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P80319, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam. Immunohistochemical analysis of paraffin-embedded human breast cancer tissue using SIRT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P80319, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam. Immunohistochemical analysis of paraffin-embedded human testis tissue using SIRT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P80319, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam. Immunohistochemical analysis of paraffin-embedded human lung cancer tissue using SIRT1 antibody. The section was pre-treated using heat mediated antigen retrieval with Tris/EDTA buffer (pH 9.0) for 20 minutes. The tissues were probed with the primary antibody (HY-P80319, 1/100) overnight at 4℃. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with neutral balsam. Immunohistochemical analysis of paraffin-embedded mouse testis tissue using SIRT1 Antibody. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody at 1/100 dilution in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX. |
| ICC | Immunocytochemistry analysis of Hela cells labeling SIRT1 with SIRT1 Antibody ( HY-P80319 ) at 1/100 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with SIRT1 Antibody ( HY-P80319 ) at 1/100 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green） was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). Immunocytochemistry analysis of Hela cells labeling SIRT1 with SIRT1 Antibody ( HY-P80319 ) at 1/200 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 15 minutes at room temperature, then blocked with quick block buffer for 10 minutes at room temperature. Cells were then incubated with SIRT1 Antibody ( HY-P80319 ) at 1/200 dilution in quick block buffer overnight at 4 ℃. AF488-conjugated Goat Anti-Rabbit IgG H&L（HY-P8002, Green） was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue). |
详细信息
背景信息
功能： SIRT1 (Sirtuin 1) 是一种 NAD\\+ 依赖性去乙酰化酶，通过调控组蛋白及多种转录调节因子的乙酰化状态，在细胞代谢、应激反应、基因组稳定性和衰老过程中发挥核心作用 [1] [2] 。从机制上看，SIRT1 将营养和能量状态与转录调控程序相连接，参与线粒体生物发生、脂肪酸氧化、葡萄糖代谢、自噬以及代谢应激适应等关键生物学过程的调控 [1] [3] 。通过与多种代谢信号网络协同作用，SIRT1 有助于维持肝脏、骨骼肌、脂肪组织和心脏等器官的代谢稳态 [1] 。在疾病相关研究中，SIRT1 活性的改变与衰老、慢性炎症状态、肥胖、代谢异常、心血管疾病、神经退行性疾病以及肿瘤相关过程密切相关，因此成为转化医学研究中广泛应用的实验靶点 [2] [4] [5] 。与其他 sirtuin 家族成员相比，SIRT1 是研究最为深入的哺乳动物 sirtuin 亚型，其主要通过去乙酰化细胞核内组蛋白和非组蛋白底物介导转录调控，而其他亚型则具有不同的亚细胞定位和底物偏好性 [1] [2] [4] 。在实验应用方面，SIRT1 可通过小分子激动剂或抑制剂进行调控；白藜芦醇 (resveratrol) 等天然多酚已被报道能够在多种模型中激活 SIRT1，因此常被用于代谢、炎症、氧化应激及衰老机制研究 [3] [4] 。 亚细胞定位： 细胞核，PML 体；细胞质；细胞核；细胞质；线粒体 表达水平： 组织特异性：广泛表达 诱导：甲基磺酸甲酯 (MMS) 可上调其表达。在 H293T 细胞中，大鼠热量限制 (CR) 血清的存在也会上调其表达。 阳性样本：HeLa cell lysate, HEK-293 cell lysate, A549 cell lysate, F9 cell lysate, Mouse testis tissue lysate, Rat testis tissue lysate, Human testis tissue. 异构体 & 翻译后修饰： Human (Q96EB6) 有两种异构体：Q96EB6-1：81,681 Da（预测值）；Q96EB6-2：61,066 Da（预测值）。 多个赖氨酸残基发生甲基化。 磷酸化。 经 TNF-α 处理后，被组织蛋白酶 B 水解，产生催化失活但稳定的 75 kDa SirtT1 片段 (75SirT1)。 被 GAPDH 进行 S-亚硝基化。 多个赖氨酸残基发生乙酰化。 泛素化。 亚基： 与 XBP1 亚型 2 相互作用 (PubMed：20955178)。与 PCAF 和 MYOD1 形成复合物。与 FOXO1 相互作用；该相互作用使 FOXO1 去乙酰化，导致其滞留在细胞核内并促进其转录活性。eNoSC 复合物的组成部分，该复合物由 SIRT1、SUV39H1 和 RRP8 组成。与 HES1、HEY2 和 PML 相互作用。与 RPS19BP1/AROS 相互作用。与 CCAR2 (通过 N 端) 相互作用；该相互作用破坏 SIRT1 与 p53/TP53 之间的相互作用。与 SETD7 相互作用；该相互作用诱导 SIRT1 从 p53/TP53 上解离并增强 p53/TP53 的活性。与 MYCN、NR1I2、CREBZF、TSC2、TLE1、FOS、JUN、NR0B2、PPARG、NCOR、IRS1、IRS2 和 NMNAT1 相互作用。与 HNF1A 相互作用；该相互作用发生在营养限制条件下。与 SUZ12 相互作用；该相互作用介导与 PRC4 组蛋白甲基化复合物的结合，这种结合具有特异性，因为未发现与 PCR2 和 PCR3 复合物变体的结合。与 BCL6 相互作用；导致特定靶基因的表观遗传抑制。与 CLOCK、BMAL1 和 PER2 相互作用 (基于相似性)。与 PPARA 相互作用；该相互作用似乎受 NAD (+) 水平的调节 (PubMed:24043310)。与 NR1H3 相互作用，且该相互作用在 CCAR2 存在的情况下受到抑制。与 CHEK2 相互作用。与 p53/TP53 相互作用。优先与 SUMO 化的 CCAR2 相互作用，而非未修饰的 CCAR2。与 PACS2 相互作用 (PubMed:29656858)。与 SIRT7 相互作用 (基于相似性)。与 PUS7 相互作用 (PubMed:31451225)。与 TULP3 相互作用 (PubMed:35397207)。与 MORN3 相互作用；该相互作用增强 p53/TP53 的泛素化 (PubMed:29681526)。