Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Vinculin antibody [EPR19579] (ab207440)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19579] to Vinculin
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Vinculin antibody [EPR19579]
    See all Vinculin primary antibodies

  • Description

    Rabbit monoclonal [EPR19579] to Vinculin

  • Host species

    Rabbit

  • Tested applications

    Suitable for: WBmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Vinculin aa 1-300. The exact sequence is proprietary.
    Database link: P18206

  • Positive control

    • WB: K562, HeLa, HEK-293T, HUVEC, MCF7, PC-3, C6, PC-12, A431, and NIH/3T3 whole cell lysates; human testis, fetal heart and fetal kidney lysates; mouse heart, kidney and spleen lysates; rat kidney and spleen lysates.

  • 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
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

Properties

Applications

Our Abpromise guarantee covers the use of ab207440 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
WB 1/1000. Detects a band of approximately 124 kDa (predicted molecular weight: 124 kDa).

Target

  • 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
    modifications

    Phosphorylated; 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

  • Database links

    see all

  • Alternative names

    • CMD1W antibody
    • CMH15 antibody
    • Epididymis luminal protein 114 antibody
    • HEL114 antibody
    • Metavinculin antibody
    • MV antibody
    • MVCL antibody
    • OTTHUMP00000019861 antibody
    • OTTHUMP00000019862 antibody
    • VCL antibody
    • VINC antibody
    • VINC_HUMAN antibody
    • Vinculin antibody
    see all

Images

  • Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    All lanes : Anti-Vinculin antibody [EPR19579] (ab207440) at 1/1000 dilution

    Lane 1 : Wild-type HeLa cell lysate
    Lane 2 : Vinculin knockout HeLa cell lysate
    Lane 3 : HEK-293 cell lysate
    Lane 4 : Jurkat cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 124 kDa



    Lanes 1-4: Merged signal (red and green). Green - ab207440 observed at 124 kDa. Red - loading control ab7291 observed at 50 kDa.

     ab207440 Anti-Vinculin antibody [EPR19579] was shown to specifically react with Vinculin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab265580 (knockout cell lysate ab257795) was used. Wild-type and Vinculin knockout samples were subjected to SDS-PAGE. ab207440 and Anti-alpha Tubulin antibody [DM1A] - Loading Control (ab7291) were incubated overnight at 4°C at 1 in 1000 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

     

  • Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    All lanes : Anti-Vinculin antibody [EPR19579] (ab207440) at 1/1000 dilution

    Lane 1 : Wild-type A431 whole cell lysate
    Lane 2 : VCL knockout A431 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : Jurkat whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 124 kDa
    Observed band size: 124 kDa



    Lanes 1 - 4: Merged signal (red and green). Green - ab207440 observed at 124 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

    ab207440 was shown to react with VCL in A431 wild-type cells in Western blot. Loss of signal was observed when VCL knockout sample was used. A431 wild-type and VCL knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab207440 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    All lanes : Anti-Vinculin antibody [EPR19579] (ab207440) at 1/2000 dilution

    Lane 1 : K562 (Human chronic myelogenous leukemia cell line from bone marrow ) whole cell lysate
    Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
    Lane 3 : HUVEC (Human umbilical vein endothelial cell line) whole cell lysate
    Lane 4 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
    Lane 5 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
    Lane 6 : Human testis lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 124 kDa
    Observed band size: 124 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure time: Lane 1-4: 3 minutes; Lane 5/6: 5 seconds.

  • Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    All lanes : Anti-Vinculin antibody [EPR19579] (ab207440) at 1/1000 dilution

    Lane 1 : Human fetal heart lysate
    Lane 2 : Human fetal kidney lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

    Predicted band size: 124 kDa
    Observed band size: 124 kDa


    Exposure time: 30 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    Western blot - Anti-Vinculin antibody [EPR19579] (ab207440)
    All lanes : Anti-Vinculin antibody [EPR19579] (ab207440) at 1/1000 dilution

    Lane 1 : Mouse heart lysate
    Lane 2 : Mouse kidney lysate
    Lane 3 : Mouse spleen lysate
    Lane 4 : Rat kidney lysate
    Lane 5 : Rat spleen lysate
    Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
    Lane 7 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate
    Lane 8 : NIH/3T3 (Mouse embryonic fibroblast cell line) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

    Predicted band size: 124 kDa
    Observed band size: 124 kDa



    Blocking/Dilution buffer: 5% NFDM/TBST.

    Exposure times: Lane 1-3: 30 seconds; Lane 4-8: 10 seconds.

References (1)

Publishing research using ab207440? Please let us know so that we can cite the reference in this datasheet.

ab207440 has been referenced in 1 publication.

  • Majmudar JD  et al. 4'-Phosphopantetheine and long acyl chain-dependent interactions are integral to human mitochondrial acyl carrier protein function. Medchemcomm 10:209-220 (2019). PubMed: 30881609

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