Anti-Vinculin antibody (ab73412)

Overview

  • Product nameAnti-Vinculin antibody
    See all Vinculin primary antibodies
  • Description
    Rabbit polyclonal to Vinculin
  • Tested applicationsSuitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Human
    Predicted to work with: Horse, Chicken, Cow, Pig, Chimpanzee, Zebrafish, Rhesus monkey, Zebra finch
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 1100 to the C-terminus of Human Vinculin.

    (Peptide available as ab74076.)

  • Positive control
    • WB: Mouse kidney, human small intestine, human skeletal muscle, rat liver and mouse pancreas tissue lysates and EBS E14 TG2A Day 3 and HepG2 whole cell lysates. ICC/IF: A549 cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab73412 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 Use a concentration of 1 µg/ml. Detects a band of approximately 130 kDa (predicted molecular weight: 124 kDa).
ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use at an assay dependent concentration.

ab171870 - Rabbit polyclonal IgG, is suitable for use as an isotype control with this antibody.

Target

  • FunctionActin 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 specificityMetavinculin is muscle-specific.
  • Involvement in diseaseDefects 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 similaritiesBelongs to the vinculin/alpha-catenin family.
  • DomainExists 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 localizationCytoplasm > 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
  • 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

Anti-Vinculin antibody images

  • ICC/IF image of ab73412 stained A549 cells. The cells were 4% formaldehyde fixed (10 min) then permeabilised using 0.1% PBS-Triton and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to further permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab73412 at 1µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

  • All lanes : Anti-Vinculin antibody (ab73412) at 1 µg/ml

    Lane 1 : Kidney (Mouse) Tissue Lysate
    Lane 2 : EBS E14 TG2A Day 3 (Mouse Pluripotent Embryonic Stem Cell) Whole Cell Lysate
    Lane 3 : HepG2 (Human hepatocellular liver carcinoma cell line) Whole Cell Lysate
    Lane 4 : Human small intestine tissue lysate - total protein (ab29276)
    Lane 5 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 6 : Liver (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    Goat polyclonal to Rabbit IgG - H&L - Pre-Adsorbed (HRP) at 1/3000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 124 kDa
    Observed band size : 130 kDa (why is the actual band size different from the predicted?)


    Exposure time : 2 minutes
  • ab73412 staining vinculin in Human platelet cells by Flow cytometry.
    Cells were fixed in paraformaldehyde and permeabilized using 0.1% Triton-X-100 in 2% BSA for 15 minutes. Primary antibody used at a 1/250 dilution and incubated for 16 hours at 4°C. The secondary antibody used was an Alexa Fluor®488 conjugated chicken anti-rabbit IgG (H+L) at a 1/500 dilution.

    P : Permeabilized
    US : Unstained (Red Peak)
    IGG RB : IgG Rabbit (Blue Peak)
    VINCULIN Ab (Green Peak)

    See Abreview

  • Anti-Vinculin antibody (ab73412) at 1/1500 dilution + whole cell lysate prepared from human platelets (treated with ADP for 30 minutes) at 20 µg

    Secondary
    HRP conjugated goat anti-rabbit polyclonal at 1/10000 dilution
    Developed using the ECL technique

    Predicted band size : 124 kDa


    Exposure time : 1 minute

    Image courtesy of Dr Mahesh Shivananjappa by Abreview.

    Blocked with 5% milk for 1 hour at 22°C.

    See Abreview

  • Immunofluorescence analysis of porcine Valvular Interstitial Cells (VIC), staining Vinculin (red) with ab73412.

    Cells were fixed with paraformaldehyde and permeabilized with 0.1% Tween 20. Samples were blocked and incubated with primary antibody (1/400 in 1% BSA) for 2 hours at 25°C. An AlexaFluor®647-conjugated donkey anti-rabbit polyclonal IgG (1/400) was used as the secondary antibody.

    See Abreview

References for Anti-Vinculin antibody (ab73412)

ab73412 has not yet been referenced specifically in any publications.

Product Wall

Application Western blot
Sample Pig Tissue lysate - whole (endometrial tissue lysate)
Gel Running Conditions Reduced Denaturing (12%)
Loading amount 10 µg
Specification endometrial tissue lysate
Blocking step Milk as blocking agent for 1 hour(s) and 30 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Feb 12 2016

Application Immunocytochemistry
Blocking step BSA as blocking agent for 30 minute(s) · Concentration: 2% · Temperature: 24°C
Sample Mouse Cell (RAW264.7)
Specification RAW264.7
Permeabilization Yes - 0.1% Triton + 2% BSA in PBS
Fixative Paraformaldehyde
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Jan 13 2014

Application Western blot
Loading amount 10 µg
Gel Running Conditions Reduced Denaturing (10%)
Sample Sheep Tissue lysate - other (Cardiac)
Specification Cardiac
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Abcam user community

Verified customer

Submitted Dec 18 2013

Application Immunocytochemistry/ Immunofluorescence
Sample Pig Cell (Valvular Interstitial Cell (VIC))
Specification Valvular Interstitial Cell (VIC)
Fixative Paraformaldehyde
Permeabilization Yes - .1% Tween20
Blocking step No blocking step used for 2 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 25°C
Username

Mr. Matthew Rush

Verified customer

Submitted Oct 22 2012

I am very pleased to hear you would like to accept our offer and test ab73412 in pig. This code will give you: 1 free PRIMARY ANTIBODY before the expiration date. To redeem this offer, please submit an Abreview for pig and include this code in the “Add...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Platelets)
Loading amount 20 µg
Specification Platelets
Treatment ADP for 30 min
Gel Running Conditions Reduced Denaturing
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
Username

Dr. Mahesh Shivananjappa

Verified customer

Submitted Nov 23 2011

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Flow Cytometry
Sample Human Cell (Platelets)
Specification Platelets
Preparation Cell harvesting/tissue preparation method: PL were isolated spinning Platelet rich plasma on Histopaque
Sample buffer: PBS
Fixation Paraformaldehyde
Permeabilization Yes - 0.1% Triton-X100 in 2% BSA for 15min
Gating Strategy Platelets
Username

Abcam user community

Verified customer

Submitted Dec 21 2010

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"