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)) 、NIH-3T3 (lane 3(20μg)) and C6 (lane 4(20μg)) using Vinculin Antibody ( HY-P80372 ). 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 (β Tubulin, 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 WT Hela cells (lane 1(20μg)) and KD Hela cells (lane 2(20μg)) using Vinculin Antibody ( HY-P80372 ). 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 ( Hsp90, 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 HepG2 (lane 2(20μg), Hela(lane 3(20μg) and NIH3T3(lane 4(20μg) using Vinculin( HY-P80372 ) 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-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature. |
| IHC-P | Immunohistochemical analysis of paraffin-embedded Mouse testis tissue using Vinculin Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) 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 (HY-P80372, 1/1000) 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 HUVEC cells labeling Vinculin with Vinculin Antibody ( HY-P80372 )at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Vinculin Antibody ( HY-P80372 ) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure 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 Vinculin with Vinculin Antibody ( HY-P80372 ) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Vinculin Antibody ( HY-P80372 ) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure 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). |
推荐稀释比例
- WB1:500-1:2000
- ICC/IF1:50-1:200
- IPUse at an assay dependent concentration.
- IHC-P1:200
详细信息
免疫原Synthetic peptide corresponding to Human Vinculin.AA range:1000-1060.
宿主/亚型Rabbit / IgG
抗体类型Recombinant,Monoclonal
全称Vinculin 抗体 (YA010)
理论分子量124 kDa
实际/观测分子量124 kDa 请注意：因蛋白存在修饰或聚体等情况，以实测为准，预测仅为参考。
产品形式溶液
偶联物Non-conjugated
储存缓冲液Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.
储存条件保存条件 & 期限: Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.
运输条件: Shipping with blue ice.
纯化方式Protein A affinity purified.
Gene ID7414 [ NCBI ]
RRIDAB_3102462
UniProt IDP18206
背景信息
功能： 纽蛋白是一种肌动蛋白丝 (F-肌动蛋白) 结合蛋白，参与细胞-基质黏附和细胞-细胞黏附，调节细胞表面 E-钙黏蛋白的表达，并增强 E-钙黏蛋白复合物的机械传感功能。它还可能在细胞形态和运动中发挥重要作用。 亚细胞定位： 细胞膜；外周膜蛋白；胞质侧；细胞连接，黏着连接；细胞连接，局灶黏着；胞质，细胞骨架；细胞膜，肌膜；外周膜蛋白；胞质侧；细胞突起，足突 表达水平： 组织特异性：Metavinculin 是一种肌肉特异性蛋白。 亚基： 具有自缔合特性。是 THSD1、PTK2/FAK1、TLN1 和 VCL 组成的复合物的一部分 (PubMed:29069646)。