Anti-Vinculin (phospho Y822) antibody (ab200825)
Key features and details
- Rabbit polyclonal to Vinculin (phospho Y822)
- Suitable for: IHC-P, WB
- Reacts with: Mouse, Rat, Recombinant fragment
- Isotype: IgG
Overview
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Product name
Anti-Vinculin (phospho Y822) antibody
See all Vinculin primary antibodies -
Description
Rabbit polyclonal to Vinculin (phospho Y822) -
Host species
Rabbit -
Tested applications
Suitable for: IHC-P, WBmore details -
Species reactivity
Reacts with: Mouse, Rat, Recombinant fragment
Predicted to work with: Human -
Immunogen
Synthetic peptide corresponding to Human Vinculin (phospho Y822). chemically synthesized phosphopeptide derived from a region of human vinculin that contains tyrosine 822.
Database link: P18206-1 -
Positive control
- WB: Chick Embryo Fibroblasts (CEFs) transfected with activated Src; IHC-P: Mouse and rat heart tissues.
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General notes
The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. Please check that this product meets your needs before purchasing.
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Properties
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Form
Liquid -
Storage instructions
Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.30
Preservative: 0.05% Sodium azide
Constituents: 50% Glycerol (glycerin, glycerine), 0.1% BSA, 49% PBS -
Concentration information loading...
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Purity
Immunogen affinity purified -
Purification notes
ab200825 has been negatively preadsorbed using a non-phosphopeptide corresponding to the site of phosphorylation to remove antibody that is reactive with non-phosphorylated vinculin. The final product is generated by affinity chromatography using a vinculin-derived peptide that is phosphorylated at tyrosine 822. -
Clonality
Polyclonal -
Isotype
IgG -
Research areas
Associated products
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Compatible Secondaries
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Isotype control
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Recombinant Protein
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab200825 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-P |
1/10 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Predicted molecular weight: 124 kDa.
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Notes |
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IHC-P
1/10 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
WB
1/1000. Predicted molecular weight: 124 kDa. |
Target
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Function
Actin filament (F-actin)-binding protein involved in cell-matrix adhesion and cell-cell adhesion. Regulates cell-surface E-cadherin expression and potentiates mechanosensing by the E-cadherin complex. May also play important roles in cell morphology and locomotion. -
Tissue specificity
Metavinculin is muscle-specific. -
Involvement in disease
Defects in VCL are the cause of cardiomyopathy dilated type 1W (CMD1W) [MIM:611407]. Dilated cardiomyopathy is a disorder characterized by ventricular dilation and impaired systolic function, resulting in congestive heart failure and arrhythmia. Patients are at risk of premature death.
Defects in VCL are the cause of cardiomyopathy familial hypertrophic type 15 (CMH15) [MIM:613255]. It is a hereditary heart disorder characterized by ventricular hypertrophy, which is usually asymmetric and often involves the interventricular septum. The symptoms include dyspnea, syncope, collapse, palpitations, and chest pain. They can be readily provoked by exercise. The disorder has inter- and intrafamilial variability ranging from benign to malignant forms with high risk of cardiac failure and sudden cardiac death. -
Sequence similarities
Belongs to the vinculin/alpha-catenin family. -
Domain
Exists in at least two conformations. When in the closed, 'inactive' conformation, extensive interactions between the head and tail domains prevent detectable binding to most of its ligands. It takes on an 'active' conformation after cooperative and simultaneous binding of two different ligands. This activation involves displacement of the head-tail interactions and leads to a significant accumulation of ternary complexes. The active form then binds a number of proteins that have both signaling and structural roles that are essential for cell adhesion.
The N-terminal globular head (Vh) comprises of subdomains D1-D4. The C-terminal tail (Vt) binds F-actin and cross-links actin filaments into bundles. An intramolecular interaction between Vh and Vt masks the F-actin-binding domain located in Vt. The binding of talin and alpha-actinin to the D1 subdomain of vinculin induces a helical bundle conversion of this subdomain, leading to the disruption of the intramolecular interaction and the exposure of the cryptic F-actin-binding domain of Vt. Vt inhibits actin filament barbed end elongation without affecting the critical concentration of actin assembly. -
Post-translational
modificationsPhosphorylated; on serines, threonines and tyrosines. Phosphorylation on Tyr-1133 in activated platelets affects head-tail interactions and cell spreading but has no effect on actin binding nor on localization to focal adhesion plaques.
Aceylated; mainly by myristic acid but also small amount of palmitic acid. -
Cellular localization
Cytoplasm > cytoskeleton. Cell junction > adherens junction. Cell membrane. Cytoplasmic face of adhesion plaques. Recruitment to cell-cell junctions occurs in a myosin II-dependent manner. Interaction with CTNNB1 is necessary for its localization to the cell-cell junctions. - Information by UniProt
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Database links
- Entrez Gene: 7414 Human
- Entrez Gene: 22330 Mouse
- Entrez Gene: 305679 Rat
- Omim: 193065 Human
- SwissProt: P18206 Human
- SwissProt: Q64727 Mouse
- SwissProt: P85972 Rat
- Unigene: 643896 Human
see all -
Alternative names
- CMD1W antibody
- CMH15 antibody
- Epididymis luminal protein 114 antibody
see all
Images
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All lanes : Anti-Vinculin (phospho Y822) antibody (ab200825) at 1/1000 dilution
Lane 1 : Untransfected CEF lysates
Lane 2 : Src transfected CEF lysates
Lane 3 : Src transfected CEF lysates with non-phosphopeptide corresponding to immunogen
Lane 4 : Src transfected CEF lysates with generic phosphotyrosine containing peptide
Lane 5 : Src transfected CEF lysates with phosphopeptide immunogen
Secondary
Lanes 1-2 : goat F(ab’)2 anti-rabbit IgG HRP conjugate
Developed using the ECL technique.
Predicted band size: 124 kDa10% polyacrylamide gel transferred to PVDF blocked with a 5% BSA-TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of rat heart tissue labeling Vinculin (phospho Y822) with ab200825 (right). Heat mediated 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 ab200825 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. Negative control without primary antibody (left).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of mouse heart tissue labeling Vinculin (phospho Y822) with ab200825 (right). Heat mediated 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 ab200825 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. Negative control without primary antibody (left).
Datasheets and documents
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SDS download
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Datasheet download
References (1)
ab200825 has been referenced in 1 publication.
- Rajakylä EK et al. Assembly of Peripheral Actomyosin Bundles in Epithelial Cells Is Dependent on the CaMKK2/AMPK Pathway. Cell Rep 30:4266-4280.e4 (2020). PubMed: 32209483