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Immunofluorescence analysis of Vinculin was done on 70% confluent log phase HeLa cells. The cells were fixed with 4% paraformaldehyde for 15 minutes; permeabilized with 0.25% Triton™ X-100 for 10 minutes followed by blocking with 5% BSA for 1 hour at room temperature. The cells were incubated with ABfinity™ Vinculin Recombinant Rabbit Monoclonal Antibody (700062) at 2 µg-4 µg in 1% BSA and incubated for 3 hours at room temperature and then labeled with Alexa Flour® 488 Goat Anti-Rabbit IgG Secondary Antibody (A11008) at a dilution of 1:400 for 30 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 594 Phalloidin (A12381). Panel d is a merged image showing junction and cytoplasmic localization of Vinculin. Panel e shows no primary antibody control. The images were captured at 20X magnification.
|Tested species reactivity||Human, Mouse|
|Published species reactivity||Primate, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||A recombinant protein corresponding to amino acids 408-591 of P18206.|
|Contains||0.09% sodium azide|
|Storage Conditions||Maintain refrigerated at 2-8°C for up to 1 month. For long term storage store at -20°C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||2-4µg/10^6 cells|
|Western Blot (WB)||1-3µg/ml|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
This antibody is predicted to react with chimpanzee, Rhesus monkey, porcine, equine, rat, bovine and chicken based on sequence homology.
ABfinity™ recombinant antibodies are rabbit monoclonal antibodies, unmatched for producing superior results. ABfinity™ antibodies are developed by immunizing animals, screening for functionality, cloning the immunogen-specific antibody genes into high-level mammalian expression vectors, produced on a large scale, and purified with Protein A.
ABfinity™ monoclonal antibodies resemble rabbit monoclonals isolated from serum or produced by hybridomas, but demonstrate greater specificity and sensitivity. Because ABfinity™ recombinant antibodies are derived from cloned DNA sequences of the heavy and light antibody chains, they are not susceptible to cell-line drift or lot-to-lot variation, thus allowing for peak specificity and performance.
Intact IgG appears on a non-reducing gel as ~150 kDa band and upon reduction generating a ~25 kDa light chain band and a ~50 kDa heavy chain.
Vinculin (130 kDa) is a ubiquitously expressed cytoskeletal protein that is involved in cell adhesion and migration and plays an important role in embryogenesis. Vinculin is phosphorylated at several residues sites within the tail domain. The vinculin protein consists of a globular head domain connected to an elongated tail region by a proline-rich domain. The head region contains binding sites for two cytoskeletal proteins, alpha-actinin and talin, as well as a binding site for the tail region of vinculin itself. The tail region contains binding sites for actin and paxillin. Vinculin head controls integrin dynamics whereas the tail controls the functional link between the focal adhesions and the actin cytoskeleton. Cell adhesion and migration require the coordinated formation and release of focal adhesions. During this process, the exchangeable vinculin fraction rises during movement and declines upon maturation.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
A multi-omics approach identifies key hubs associated with cell type-specific responses of airway epithelial cells to staphylococcal alpha-toxin.
700062 was used in immunocytochemistry to use a multi-omics strategy to elucidate the cellular programs altered by Staphylococcus aureus alpha-toxin
|Richter E,Harms M,Ventz K,Gierok P,Chilukoti RK,Hildebrandt JP,Mostertz J,Hochgräfe F||PloS one (10:null)||2015|
Force engages vinculin and promotes tumor progression by enhancing PI3K activation of phosphatidylinositol (3,4,5)-triphosphate.
700062 was used in immunocytochemistry to study the mechanism of the activation of focal adhesion protein vinculin by mechanical stress
|Rubashkin MG,Cassereau L,Bainer R,DuFort CC,Yui Y,Ou G,Paszek MJ,Davidson MW,Chen YY,Weaver VM||Cancer research (74:4597)||2014|
Dual-objective STORM reveals three-dimensional filament organization in the actin cytoskeleton.
700062 was used in immunocytochemistry to develop an astigmatism imaging/dual objective STORM system for biological imaging using the actin cytoskeleton as a model system
|Xu K,Babcock HP,Zhuang X||Nature methods (9:185)||2012|
CMD1W; CMH15; epididymis luminal protein 114; metavinculin; MVCL; vinculin
9430097D22; AA571387; AI462105; AW545629; CMD1W; CMH15; HEL114; MV; MVCL; VCL