Recombinant Anti-Vinculin antibody [EPR8185] (ab129002)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR8185] to Vinculin
- Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
- Reacts with: Mouse, Rat, Human
Related conjugates and formulations
Overview
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Product name
Anti-Vinculin antibody [EPR8185]
See all Vinculin primary antibodies -
Description
Rabbit monoclonal [EPR8185] to Vinculin -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IP, ICC/IFmore details
Unsuitable for: IHC-P -
Species reactivity
Reacts with: Mouse, Rat, Human -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
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Positive control
- WB: PC3, HeLa, U937, K562, HUVEC, HepG2 human fetal liver and human fetal kidney lysates ICC/IF: HUVEC cells, HEK-293 cells. IP: HeLa cells Flow Cyt (intra): HEK293 cells, A-431 cells.
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General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.
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. Stable for 12 months at -20°C. -
Storage buffer
pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR8185 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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Isotype control
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab129002 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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Flow Cyt (Intra) |
1/200.
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WB | (16) |
1/10000 - 1/50000. Detects a band of approximately 124 kDa (predicted molecular weight: 124 kDa).
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IP |
1/10 - 1/100.
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ICC/IF | (2) |
1/50 - 1/250.
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Notes |
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Flow Cyt (Intra)
1/200. |
WB
1/10000 - 1/50000. Detects a band of approximately 124 kDa (predicted molecular weight: 124 kDa). |
IP
1/10 - 1/100. |
ICC/IF
1/50 - 1/250. |
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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Flow cytometry overlay histogram showing wild-type A-431 (green line) and VCL knockout A-431 stained with ab129002 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilised with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS containing 10% normal goat serum to block non-specific protein-protein interaction followed by the antibody (ab129002) (1x 106 in 100μl at 0.00032 μg/ml (1/6249999)) for 30min at 22°C.
The secondary antibody Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) preadsorbed was incubated at 1/4000 for 30min at 22°C
Isotype control antibody Recombinant Rabbit IgG, monoclonal [EPR25A] - Isotype Control was used at the same concentration and conditions as the primary antibody (wild-type A-431 - black line, VCL knockout A-431 - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).
Acquisition of >5000 events were collected using a 50 mW Blue laser (488nm) and 525/40 bandpass filter.
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Immunofluorescent staining of HEK293 cells (fixed in 4% PFA, permeabilized with 0.1% Triton X 100) using purified ab129002 at a dilution of 1/50. An Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) was used as the secondary at a dilution of 1/1000 and the cells were counter stained with DAPI. The negative controls are shown in the bottom middle and right hand panels. For negative control 1, the primary was used and then goat anti-mouse IgG was used at a dilution of 1/500. For negative control 2, a mouse primary antibody (ab7291) and anti-rabbit secondary antibody (ab150077) were used.
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All lanes : Anti-Vinculin antibody [EPR8185] (ab129002) at 1/10000 dilution (purified)
Lane 1 : HepG2 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : PC3 cell lysate
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 124 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Intracellular Flow Cytometry analysis of 293 (human embryonic kidney epithelial) cells labeling Vinculin (red) with ab129002 at a 1/200 dilution. Cells were fixed with 4% paraformaldehyde and permeabilized with 90% methanol. A goat anti-rabbit IgG (Alexa Fluorr® 488) (ab150077) was used as the secondary antibody at a 1/2000 dilution. Black - Rabbit monoclonal IgG (ab172730). Blue (unlabeled control) - Cells without incubation with the primary and secondary antibodies.
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ab129002 (purified) at 1/20 immunoprecipitating vinculin in HeLa cells. Lane 1: HeLa whole cell lysate (10 µg). Lane 2: HeLa whole cell lysate (10 µg). Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab129002 in HeLa whole cell lysate. For western blotting, a HRP-conjugated goat anti-rabbit antibody was used as the secondary antibody (1/1000).
Blocking buffer and concentration: 5% NFDM/TBST.
Diluting buffer and concentration: 5% NFDM /TBST.
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Different batches of ab129002 were tested on HepG2 (Human hepatocellular carcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 124 kDa.
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All lanes : Anti-Vinculin antibody [EPR8185] (ab129002) at 1/20000 dilution (purified)
Lane 1 : C6 cell lysate
Lane 2 : rat kidney cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 124 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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Immunofluorescent staining of vinculin in HUVEC cells with unpurified ab129002 at 1/100 dilution.
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All lanes : Anti-Vinculin antibody [EPR8185] (ab129002) at 1/10000 dilution (purified)
Lane 1 : mouse spleen
Lane 2 : NIH/3T3 cell lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : HRP goat anti-rabbit (H+L) at 1/1000 dilution
Predicted band size: 124 kDa
Observed band size: 124 kDaBlocking buffer: 5% NFDM/TBST
Dilution buffer: 5% NFDM/TBST
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All lanes : Anti-Vinculin antibody [EPR8185] (ab129002) at 1/10000 dilution (unpurified)
Lane 1 : PC3 cell lysate
Lane 2 : HeLa cell lysate
Lane 3 : U937 cell lysate
Lane 4 : K562 cell lysate
Lane 5 : HUVEC cell lysate
Lane 6 : Human fetal liver lysate
Lane 7 : Human fetal kidney lysate
Lysates/proteins at 10 µg per lane.
Secondary
All lanes : Goat anti-Rabbit HRP at 1/2000 dilution
Predicted band size: 124 kDa
Observed band size: 124 kDa
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (277)
ab129002 has been referenced in 277 publications.
- Hühn D et al. Prolonged estrogen deprivation triggers a broad immunosuppressive phenotype in breast cancer cells. Mol Oncol 16:148-165 (2022). PubMed: 34392603
- Abd-Elrahman KS et al. A positive allosteric modulator for the muscarinic receptor (M1 mAChR) improves pathology and cognitive deficits in female APPswe/PSEN1ΔE9 mice. Br J Pharmacol 179:1769-1783 (2022). PubMed: 34820835
- Drazic A et al. The Final Maturation State of β-actin Involves N-terminal Acetylation by NAA80, not N-terminal Arginylation by ATE1. J Mol Biol 434:167397 (2022). PubMed: 34896361
- Cheng B et al. Anti-PD-L1/TGF-βR fusion protein (SHR-1701) overcomes disrupted lymphocyte recovery-induced resistance to PD-1/PD-L1 inhibitors in lung cancer. Cancer Commun (Lond) 42:17-36 (2022). PubMed: 34981670
- Duran I et al. 4-PBA Treatment Improves Bone Phenotypes in the Aga2 Mouse Model of Osteogenesis Imperfecta. J Bone Miner Res 37:675-686 (2022). PubMed: 34997935