Anti-Vinculin antibody [VIN-54] (ab130007)

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Western blot - Anti-Vinculin antibody [VIN-54] (ab130007)
  • Immunohistochemistry (Frozen sections) - Anti-Vinculin antibody [VIN-54] (ab130007)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [VIN-54] (ab130007)
  • Immunohistochemistry (Frozen sections) - Anti-Vinculin antibody [VIN-54] (ab130007)
  • Western blot - Anti-Vinculin antibody [VIN-54] (ab130007)

Key features and details

  • Mouse monoclonal [VIN-54] to Vinculin
  • Suitable for: ICC/IF, WB, IHC-Fr
  • Reacts with: Mouse, Rat, Human, African green monkey
  • Isotype: IgG1

Overview

  • Product name

    Anti-Vinculin antibody [VIN-54]
    See all Vinculin primary antibodies

  • Description

    Mouse monoclonal [VIN-54] to Vinculin

  • Host species

    Mouse

  • Tested applications

    Suitable for: ICC/IF, WB, IHC-Frmore details

  • Species reactivity

    Reacts with: Mouse, Rat, Human, African green monkey

  • Immunogen

    Human Vinculin purified from uterus.

  • Positive control

    • WB: Hela, HepG2, Cos-7, U-87 MG, PC-12, RH35, HEPA1-6 and NIH-3T3 whole cell lysates; ICC/IF: Human mammary cancer cells; IHC-P: Human breast tissue.

  • 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.

    If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, along with publications, customer reviews and Q&As

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab130007 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
ICC/IF
Use a concentration of 2 µg/ml.
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 124 kDa.
IHC-Fr
Use a concentration of 2 - 4 µg/ml.
Notes
ICC/IF
Use a concentration of 2 µg/ml.
WB
Use a concentration of 1 - 2 µg/ml. Predicted molecular weight: 124 kDa.
IHC-Fr
Use a concentration of 2 - 4 µg/ml.

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 [VIN-54] (ab130007)
    Western blot - Anti-Vinculin antibody [VIN-54] (ab130007)
    All lanes : Anti-Vinculin antibody [VIN-54] (ab130007) at 0.5 µg/ml

    Lane 1 : Hela whole cell lysates
    Lane 2 : HepG2 whole cell lysates
    Lane 3 : Monkey COS-7 whole cell lysates
    Lane 4 : U-87MG whole cell lysates
    Lane 5 : Rat PC-12 whole cell lysates
    Lane 6 : Rat RH35 whole cell lysates
    Lane 7 : Mouse HEPA1-6 whole cell lysates
    Lane 8 : Mouse NIH3T3 whole cell lysates

    Lysates/proteins at 50 µg per lane.

    Secondary
    All lanes : Goat anti- mouse IgG-HRP at 1/10000 dilution

    Developed using the ECL technique.

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



    The signal is developed using an Enhanced Chemiluminescent detection (ECL) kit with Tanon 5200 system.

    After Electrophoresis, proteins were transferred to a Nitrocellulose membrane at 150mA for 50-90 minutes. Blocked the membrane with 5% Non-fat Milk/ TBS for 1.5 hour at RT.

     

     

     

  • Immunohistochemistry (Frozen sections) - Anti-Vinculin antibody [VIN-54] (ab130007)
    Immunohistochemistry (Frozen sections) - Anti-Vinculin antibody [VIN-54] (ab130007)

    Immunohistochemical analysis of frozen sections of human placenta tissue labelling Vinculin with ab130007 at 1μg/ml, followed by a biotinylated goat anti-mouse IgG secondary antibody. The tissue was blocked with 10% goat serum. The tissue section was developed using Strepavidin-Biotin-Complex with DAB as the chromogen.

  • Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [VIN-54] (ab130007)
    Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [VIN-54] (ab130007)

    Immunocytochemistry/Immunofluorescent analysis of human mammary cancer cells labelling Vinculin with ab130007 at 2μg/ml, followed by a DyLight®488 Conjugated Goat Anti-Mouse secondary antibody at 1/100 dilution. DAPI was used as a nuclear counterstain.

     

  • Immunohistochemistry (Frozen sections) - Anti-Vinculin antibody [VIN-54] (ab130007)
    Immunohistochemistry (Frozen sections) - Anti-Vinculin antibody [VIN-54] (ab130007)

    Immunohistochemical analysis of frozen sections of human placenta tissue labelling Vinculin with ab130007 at 1μg/ml, followed by a biotinylated goat anti-mouse IgG secondary antibody. The tissue was blocked with 10% goat serum. The tissue section was developed using Strepavidin-Biotin-Complex with DAB as the chromogen.

  • Western blot - Anti-Vinculin antibody [VIN-54] (ab130007)
    Western blot - Anti-Vinculin antibody [VIN-54] (ab130007)
    All lanes : Anti-Vinculin antibody [VIN-54] (ab130007) at 1/10000 dilution

    Lane 1 : HAP1
    Lane 2 : HEK293
    Lane 3 : U-2 OS

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 124 kDa



    ab130007 was shown to recognize vinculin when whole cell lysates were subjected to SDS-PAGE. The blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes which was incubated overnight at 4°C at 0.1 μg antibody per mL and developed with Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibody at 1/15000 dilution for 1 hour at room temperature before imaging.

References (6)

Publishing research using ab130007? Please let us know so that we can cite the reference in this datasheet.

ab130007 has been referenced in 6 publications.

  • Pillai-Kastoori L  et al. Antibody validation for Western blot: By the user, for the user. J Biol Chem 295:926-939 (2020). PubMed: 31819006
  • Ferrante F  et al. HDAC3 functions as a positive regulator in Notch signal transduction. Nucleic Acids Res 48:3496-3512 (2020). PubMed: 32107550
  • Demircioglu FE  et al. The AAA?+?ATPase TorsinA polymerizes into hollow helical tubes with 8.5 subunits per turn. Nat Commun 10:3262 (2019). PubMed: 31332180
  • Simchovitz A  et al. NEAT1 is overexpressed in Parkinson's disease substantia nigra and confers drug-inducible neuroprotection from oxidative stress. FASEB J 33:11223-11234 (2019). PubMed: 31311324
  • Singh AN  et al. The p97-Ataxin 3 complex regulates homeostasis of the DNA damage response E3 ubiquitin ligase RNF8. EMBO J 38:e102361 (2019). PubMed: 31613024
  • Pietrangelo A & Ridgway ND Golgi localization of oxysterol binding protein-related protein 4L (ORP4L) is regulated by ligand binding. J Cell Sci 131:N/A (2018). PubMed: 29930082

Customer reviews and Q&As

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Immunocytochemistry/ Immunofluorescence abreview for Anti-Vinculin antibody [VIN-54]

 Inconclusive
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (184A1 mammary epithelial)
Permeabilization
Yes - PBS + 0.25% Triton-X
Specification
184A1 mammary epithelial
Blocking step
LI-COR Intercept Blocking Buffeer as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: RT°C
Fixative
Paraformaldehyde
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Submitted Sep 03 2020

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