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Vinculin is a cytoskeletal membrane protein involved in cell-cell and cell-matrix adhesion. It is known to play a role in the anchoring of F-actin to the membrane. Also it is thought to regulate the expression of cell surface E-cadherin. The Vinculin antibody detects endogenous levels of Vinculin protein.
1. HOW DID OUR ANTIBODY PERFORM? ANTIBODY CUSTOMER REVIEW: Vinculin Antibody (STJ96247)
2. What was used? Primary antibody: STJ96247 Vinculin Antibody Provider: St John’s Laboratory Dilution ratio: 1:100 Application: Immunohistochemistry Materials for validation: Skeletal muscle of human origin (kryo-section) Click for product data sheet PDF
3. What Was The Protocol? Treatment of Materials ? Cryo sections were cut into 6um pieces, and left overnight in -80¡ãC freezer. Protocol ? Primary antibody was incubated overnight at room temperature (23¡ãC). ? The sample was washed for 2 hours with multiple changes of PBS. ? Amplification occurred through NOVOLINK Leica microsystems, then the primary antibody was incubated for 1 hour. ? The sample was then washed again for 1 hour with at least 3 changes of PBS. ? The secondary antibody was incubated for 30 minutes and then washed for 2 hours using PBS with multiple changes. ? The sample was then stained for 2 minutes using DAB. ? The sample was counterstained for 3 seconds using Harris¡¯ haematoxylin and then washed with tap water for 2 minutes until blueing. ? The sample was then dehydrated and mounted in entellan.
4. What were the results?
5. What did the customer think? Antibody Specificity: Antibody RPPA Rating: Testing results were provided independently by A. Roos at Newcastle University. They stated ¡°Immunohistochemical investigation showed increased vinculin immunoreactivity (also at the sarcolemm) in degenerating myofibres which is in accordance with our proteomic in vitro data and with the predicted subcellular localization of this protein. Peculiar nuclear immunoreactivity for this actin filament (F-actin)-binding protein has also been described for an Abcam Vinculin antibody (Ab18058).¡±
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