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Key Specifications Table
| Species Reactivity | Key Applications | Host | Format | Antibody Type |
|---|---|---|---|---|
| B, H, M, Mk, Po, Rb | IP, WB, ICC, IH(P) | M | Purified | Monoclonal Antibody |
| Description | |
|---|---|
| Catalogue Number | MAB3574-25UG |
| Brand Family | Chemicon® |
| Trade Name |
|
| Description | Anti-Vinculin, clone VIIF9 (7F9) |
| Product Information | |
|---|---|
| Format | Purified |
| Control |
|
| Presentation | Liquid in 0.02M Phosphate buffer with 0.25M NaCl and 0.1% sodium azide. |
| Quality Level | MQ100 |
| Applications | |
|---|---|
| Application | Anti-Vinculin Antibody, clone VIIF9 (7F9) is an antibody against Vinculin for use in IP, WB, IC, IH(P). |
| Key Applications |
|
| Application Notes | Western blot. 1:1000; ~117kDa band. Suggested blocking buffer is TBS-Tween with 2% BSA. Suggested dilution buffer is TBS-Tween with 0.05% sodium azide. Preferred gel percentage is 10% and/or 4-20% (or similar) gradient gel. Immunohistochemistry on frozen and paraffin embedded tissue sections. Suggested fixation for frozen tissue sections is acetone fix for 6 minutes at room temperature. For formalin fixed paraffin embedded tissue sections: microwave in 0.01M citrate buffer (pH 6.0) for 8-10 minutes (note that all microwaves differ and adjustments may need to be made). If necessary follow with enzyme digestion (0.01% pronase for 10 mintues). Suggested blocking agent is fetal bovine serum. The antibody has also been used successfully on methyl-Carnoy fixed tissue. Immunocytochemistry (see application notes below). Immunoprecipitation. Suggested extraction buffer is 20 mM Tris-HCl, pH 7.4, 150 mM NaCl, 1% Triton X-100, 0.1% SDS, 0.5% deoxycholic acid-NaCl and 0.5 mM PMSF. Final reaction volume is 1 mL and suggested capture agent is agarose conjugated anti-mouse IgG. Note that the muscle-specific isoform, meta-vinculin (MW 150 kDa) will co-precipitate. Optimal working dilutions must be determined by the end user. APPLICATION NOTES FOR MAB3574 IMMUNOCYTOCHEMISTRY For cultured cells, use the following method: 1) wash cells twice with buffer "A" (100 mM Pipes (to make it dissolve, titrate with KOH), 0.5mM MgCl2, 0.1 mM EDTA, 0.01M EGTA (to make it dissolve, titrate with KOH), pH 6.9); 2) fix 15 min @RT (3.7% formalin, 0.2% Triton X-100, 2M Glycerol, prepared on buffer A); 3) wash 3 times with buffer "A"; 4) proceed with incubation with primary Abs, etc. |
| Biological Information | |
|---|---|
| Immunogen | Crude smooth muscle extract from normal human adult uterus. |
| Clone | VIIF9 (7F9) |
| Concentration | Please refer to lot specific datasheet. |
| Host | Mouse |
| Specificity | Vinculin. Cellular localization is Intracellular, associated with cytoskeleton. |
| Isotype | IgG1 |
| Species Reactivity |
|
| Antibody Type | Monoclonal Antibody |
| Entrez Gene Number |
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| Entrez Gene Summary | 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. Multiple alternatively spliced transcript variants have been found for this gene, but the biological validity of some variants has not been determined. Human vinculin protein exhibits a greater than 95% sequence identity to the chicken vinculin protein. |
| Gene Symbol |
|
| UniProt Number |
|
| UniProt Summary | FUNCTION: SwissProt: P18206 # Involved in cell adhesion. May be involved in the attachment of the actin-based microfilaments to the plasma membrane. May also play important roles in cell morphology and locomotion. SIZE: 1134 amino acids; 123799 Da SUBUNIT: Exhibits self-association properties. Interacts with NRAP and SORBS1 (By similarity). Interacts with TLN1. SUBCELLULAR LOCATION: Cytoplasm, cytoskeleton. Cell junction, adherens junction; Cytoplasmic side. Note=Cytoplasmic face of adhesion plaques. TISSUE SPECIFICITY: Metavinculin is muscle-specific. PTM: Phosphorylated on serines, threonines and tyrosines and acylated by myristic acid and/or palmitic acid. DISEASE:SwissProt: P18206 # Defects in VCL are a cause of hypertrophic cardiomyopathy (HCM) and dilated cardiomyopathy (DCM). SIMILARITY: SwissProt: P18206 ## Belongs to the vinculin/alpha-catenin family. |
| Product Usage Statements | |
|---|---|
| Quality Assurance | Tested |
| Usage Statement |
|
| Storage and Shipping Information | |
|---|---|
| Storage Conditions | Maintain at 2-8°C in undiluted aliquots up to 6 months after date of receipt |
| Packaging Information | |
|---|---|
| Material Size | 25 μg |