View (14) other Vinculin antibodies
Vinculin Monoclonal Antibody (VLN01)
Antibody target was verified by Relative expression to ensure the antibody binds to the antigen stated.
Catalog # MA5-11690
(USD) 349.00, 500 µL
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Immunofluorescence analysis of Vinculin was performed using 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Vinculin (VLN01) Mouse Monoclonal Antibody (Product # MA5-11690) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (Product # A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (Product # S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
Western blot analysis was performed on whole cell extracts (30 µg lysate) of HeLa (Lane 1), NIH/3T3 (Lane 2), COS-7 (Lane 3), Hep G2 (Lane 4), Caco-2 (Lane 5), HT-29 (Lane 6), T-47D (Lane 7), K-562 (Lane 8), U-2 OS (Lane 9), HEK-293 (Lane 10). The blots were probed with Anti-Vinculin Mouse Monoclonal Antibody (Product # MA5-11690, 1-3 µg/ml) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conj µgate (Product # A28177, 0.4 µg/ml, 1:2500 dilution). A 124 kDa band corresponding to Vinculin was observed across the cell lines tested. Known quantity of protein samples were electrophoresed using Novex® NuPAGE® 10 % Bis-Tris gel (Product # NP0302BOX), XCell SureLock™ Electrophoresis System (Product # EI0002) and Novex® Sharp Pre-Stained Protein Standard (Product # LC5800). Resolved proteins were then transferred onto a nitrocellulose membrane with overnight wet transfer system. The membrane was probed with the relevant primary and secondary Antibody using iBind™ Flex Western Starter Kit (Product # SLF2000S). Chemiluminescent detection was performed using Novex® ECL Chemiluminescent Substrate Reagent Kit (Product # WP20005).
Western blot analysis was performed on membrane enriched extracts (30 µg lysate) of T-47D (Lane 1),K-562 (Lane 2), L6 (Lane 3), C2C12 (Lane 4), COS-7 (Lane 5), MOLT4 (Lane 6), Ramos (Lane 7) and Jurkat (Lane 8). The blot was probed with Vinculin Mouse Monoclonal Antibody (Product # MA5-11690, 1:250 dilution) and detected by chemiluminescence using Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, HRP conjugate (Product # A28177, 0.25 µg/mL, 1:4000 dilution). A 124 kDa band corresponding to Vinculin was observed in T-47D, K-562, L6, C2C12, COS-7 and not observed in other cell lines which are documented to be Vinculin negative.
Western blot of Vinculin using Vinculin Monoclonal Antibody (Product # MA5-11690) on HeLa Cells.
Immunoprecipitation of Vinculin using Vinculin Monoclonal Antibody (Product # MA5-11690) on denatured Human LS174T Cells.
Antibody specificity was demonstrated by detection of differential basal expression of the target across cell models owing to their inherent genetic constitution. Relative expression of Vinculin was observed in T-47D, K-562, L6, C2C12, COS-7, MOLT4, Ramos and Jurkat in Western Blot using Vinculin Monoclonal Antibody (Product # MA5-11690).Vinculin is reported to be expressed in T-47D, K-562, L6, C2C12, COS-7 and not in other cell lines like MOLT4, Ramos and Jurkat.
Fig 4 EGFR Interaction with RSV F Depends on Strain-Specificity of F. 293T cells were co-transfected with either SLAM (control) or EGFR and RSV F constructs. Whole cell lysates were obtained after 24 h post-transfection and were either directly blotted ( A ) or immunoprecipitated with motavizumab mAb specific for RSV F ( B ). Whole cell lysates were probed for RSV-F, EGFR, SLAM, and vinculin expression. Proteins immunoprecipitated were probed for RSV-F and EGFR levels. The graph depicts ratios of EGFR to RSV F1 from four separate experiments combined. Error bars represent standard error of the mean (SEM). * and ** represent significant differences ( P < 0.05 and P < 0.005, respectively by ANOVA).
Immunohistochemistry (Paraffin) (IHC (P))
Western Blot (WB)
Host / Isotype
PBS, pH 7.4, with 0.2% BSA
0.09% sodium azide
Product Specific Information
MA5-11690 targets Vinculin in IF, IHC (P), IP, and WB applications and shows reactivity with Human samples.
The MA5-11690 immunogen is semi-purified vinculin.
Vinculin is an attachment protein involved in the indirect binding of intracellular actin filaments to extracellular fibronectin. Vinculin binds to actin capping protein and alpha actinin, which in turn binds to actin. The binding of fibronectin is also indirect where vinculin binds to talin, which in turn binds to the intracellular domain of the fibronectin receptor which itself is the ligand for fibronectin. Vinculin is widely distributed in tissue and is expressed where smooth muscle actin and fibroblasts attach to the extracellular matrix.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
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