Human/Mouse/Rat Vinculin Antibody

Detection of Human Vinculin by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Vinculin Monoclonal Antibody (Catalog # MAB6896) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Vinculin at approximately 124 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Vinculin in Human Uterus. Vinculin was detected in immersion fixed paraffin-embedded sections of human uterus using Mouse Anti-Human Vinculin Monoclonal Antibody (Catalog # MAB6896) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to smooth muscle cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

Detection of Human Vinculin by Simple Western^TM. Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and PC‑3 human prostate cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Vinculin at approximately 122 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse/Rat Vinculin Monoclonal Antibody (Catalog # MAB6896). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Detection of Human Vinculin by Western Blot Cr and Ni exposure increases tau levels and phosphorylation in iNeurons and RA-differentiated SH-SY5Y cells. (a,b) iPSCs carrying the R406W tau mutation and isogenic controls were differentiated into iNeurons and treated with Cr (5 µM) and Ni (800 µM) for 72 hours. The levels of total (a) and phospho-tauSer396/404 (PHF-1) (b) tau were measured by western blot. GAPDH and vinculin were used as loading controls. Images show representative immunoblots comparing tau and phospho-tau levels in control and mutant iNeurons before and after Cr and Ni treatment. Plots represents the average ± SEM of 3 independent experiments. (c,d) RA-differentiated SH-SY5Y cells were treated with Cr (2.5 µM) and Nickel (200 µM) for 24 hours before protein extraction. Images show representative immunoblots comparing the levels total tau (c) and phospho-tauSer396/404 (PHF-1) tau (d) before and after heavy metals treatment. GAPDH and vinculin were used as loading control. Plots represent the average ± SEM of 4 independent experiments. Statistical significance was determined by two‐ANOVA followed by Bonferroni’s test for multiple comparisons or one-way ANOVA using GraphPad Prism 6. p-values comparing untreated and heavy metal treated cells are included in the graphs. Statistical significance was considered when p-value ≤ 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31953414), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Vinculin by Western Blot Cr and Ni exposure increases tau levels and phosphorylation in iNeurons and RA-differentiated SH-SY5Y cells. (a,b) iPSCs carrying the R406W tau mutation and isogenic controls were differentiated into iNeurons and treated with Cr (5 µM) and Ni (800 µM) for 72 hours. The levels of total (a) and phospho-tauSer396/404 (PHF-1) (b) tau were measured by western blot. GAPDH and vinculin were used as loading controls. Images show representative immunoblots comparing tau and phospho-tau levels in control and mutant iNeurons before and after Cr and Ni treatment. Plots represents the average ± SEM of 3 independent experiments. (c,d) RA-differentiated SH-SY5Y cells were treated with Cr (2.5 µM) and Nickel (200 µM) for 24 hours before protein extraction. Images show representative immunoblots comparing the levels total tau (c) and phospho-tauSer396/404 (PHF-1) tau (d) before and after heavy metals treatment. GAPDH and vinculin were used as loading control. Plots represent the average ± SEM of 4 independent experiments. Statistical significance was determined by two‐ANOVA followed by Bonferroni’s test for multiple comparisons or one-way ANOVA using GraphPad Prism 6. p-values comparing untreated and heavy metal treated cells are included in the graphs. Statistical significance was considered when p-value ≤ 0.05. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31953414), licensed under a CC-BY license. Not internally tested by R&D Systems.

Detection of Human Vinculin by Western Blot Cr and Ni exposure induces apoptotic cell death in SH-SY5Y cells. Non-differentiated and RA-differentiated SH-SY5Y cells were treated with Cr (2.5 µM) and Ni (200 µM) for 24 hours. Whole cell lysates were collected in order to analyze by western blot the levels of cleaved caspase-3 protein (a), the ratio of anti-apoptotic protein Bcl2 to pro-apoptotic protein Bax (b) and the cleavage and activation of caspase-9 (c). Vinculin and GAPDH were used as loading controls. (a) Representative immunoblot showing the presence of the 17/19KD fragment of caspase-3 after Cr and Ni exposure. (b) Representative immunoblots showing the levels of Bcl2 and Bax proteins after heavy metal exposure. The activation of the intrinsic/mitochondrial apoptosis pathway was assessed by the decrease of the ratio of Bcl2 to full length Bax and the presence of a cleavage form of Bax protein. The plot represent the average ± SEM of 3 independent experiments. Statistical significance was determined by one-way analysis of variance (anova) followed by Bonferroni’s test for multiple comparisons using GraphPad Prism 6. p-values comparing untreated and heavy metal treated cells are included. Statistical significance was considered when p-value ≤ 0.05. (c) Immunoblot showing the presence of the fragment (43KD) of active caspase-9 after Cr and Ni exposure to confirm the activation of the intrinsic/mitochondrial pathway. All experiments were performed in triplicates. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/31953414), licensed under a CC-BY license. Not internally tested by R&D Systems.

Simple Western: Vinculin Antibody (728526) [Unconjugated] [MAB6896] - Simple Western: Vinculin Antibody (728526) [Unconjugated] [MAB6896] - Western blot analysis for expression levels of mTOR and its downstream factors in the DRG on PID 21. Phosphorylated expressions of mTOR complex 1 and 2 were decreased in the shmTOR group, and its downstream effectors were downregulated as well. Data are expressed as mean +- SEM, * p < 0.05, ** p < 0.01, **** p < 0.0001. One-way ANOVA with Tukey's multiple comparisons test. Image collected and cropped by CiteAb from the following publication (https://pubmed.ncbi.nlm.nih.gov/37958901), licensed under a CC-BY license. Not internally tested by R&D Systems.