Immunohistochemistry analysis of Phospho-Vinculin [pTyr822] showing staining in the cytoplasm of paraffin-embedded mouse heart tissue (right) compared to a negative control without primary antibody (left). To expose target proteins, antigen retrieval was performed using 10mM sodium citrate (pH 6.0), microwaved for 8-15 min. Following antigen retrieval, tissues were blocked in 3% H2O2-methanol for 15 min at room temperature, washed with ddH2O and PBS, and then probed with a Phospho-Vinculin [pTyr822] polyclonal antibody (441080G) diluted in 3% BSA-PBS at a dilution of 1:20 overnight at 4°C in a humidified chamber. Tissues were washed extensively in PBST and detection was performed using an HRP-conjugated secondary antibody followed by colorimetric detection using a DAB kit. Tissues were counterstained with hematoxylin and dehydrated with ethanol and xylene to prep for mounting.
|Tested species reactivity||Chicken, Mouse, Rat|
|Published species reactivity||Tag, Rat, Human|
|Host / Isotype||Rabbit / IgG|
|Immunogen||The antiserum was produced against a chemically synthesized phosphopeptide derived from a region of human vinculin that contains tyrosine 822. The sequence is conserved in mouse, rat, and chicken.|
|Purification||Antigen affinity chromatography|
|Storage buffer||Dulbecco's PBS, pH 7.3, with 1mg/ml BSA, 50% glycerol|
|Contains||0.05% sodium azide|
|Tested Applications||Dilution *|
|Immunohistochemistry (Paraffin) (IHC (P))||1:10-1:100|
|Western Blot (WB)||Assay Dependent|
|Western Blot (WB)||See 2 publications below|
Vinculin is a cytoskeletal protein associated with cell-cell and cell-matrix junctions, where it is thought to function as one of several interacting proteins involved in anchoring F-actin to the membrane. Defects in VCL are the cause of cardiomyopathy dilated type 1W. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Multiple alternatively spliced transcript variants have been found for this gene, but the biological validity of some variants has not been determined.
|Adhesion molecules affected by treatment of lung cancer cells with epidermal growth factor.||Fonseca FL,Azzalis LA,Feder D,Nogoceke E,Junqueira VB,Valenti VE,de Abreu LC||Lung (189:383)||2011|
|Human||Not Cited||The Helicobacter pylori CagA protein disrupts matrix adhesion of gastric epithelial cells by dephosphorylation of vinculin.||Moese S,Selbach M,Brinkmann V,Karlas A,Haimovich B,Backert S,Meyer TF||Cellular microbiology (9:1148)||2007|