Recombinant
RabMAb

Recombinant Anti-Vinculin antibody [EPR8185] (ab129002)

Rabbit recombinant monoclonal Vinculin antibody [EPR8185]. Validated in WB, IP, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human. Cited in 46 publication(s). Independently reviewed in 11 review(s).

Overview

  • 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, WB, IP, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Vinculin aa 1000-1100. The exact sequence is proprietary.
    Database link: P18206

  • 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: HEK293 cells.
  • General notes

      

    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.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
Flow Cyt 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

  • 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
  • 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

  • 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.

  • 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 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • 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 Fluor® 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.

  • 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.

  • 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 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • Immunofluorescent staining of vinculin in HUVEC cells with unpurified ab129002 at 1/100 dilution.

  • 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 kDa



    Blocking buffer: 5% NFDM/TBST

    Dilution buffer: 5% NFDM/TBST

  • 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

References

This product has been referenced in:
  • Kataoka Y  et al. Hypoxia-induced galectin-3 enhances RhoA function to activate the motility of tumor cells in non-small cell lung cancer. Oncol Rep 41:853-862 (2019). Read more (PubMed: 30535445) »
  • Pegg HJ  et al. The RUNX Transcriptional Coregulator, CBFß, Suppresses Migration of ER+ Breast Cancer Cells by Repressing ERa-Mediated Expression of the Migratory Factor TFF1. Mol Cancer Res N/A:N/A (2019). Read more (PubMed: 30655324) »
See all 53 Publications for this product

Customer reviews and Q&As

1-10 of 11 Abreviews or Q&A

Application
Western blot
Sample
Human Cell lysate - whole cell (PNT2 prostate tissue)
Gel Running Conditions
Reduced Denaturing (6)
Loading amount
20 µg
Specification
PNT2 prostate tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jan 08 2019

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (RAW 264.7)
Permeabilization
Yes - 0.2% Triton-X
Specification
RAW 264.7
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Paraformaldehyde

Anugraha Rajagopalan

Verified customer

Submitted Aug 17 2018

Application
Western blot
Sample
Zebrafish Tissue lysate - whole (Testicle)
Gel Running Conditions
Reduced Denaturing (12)
Loading amount
15 µg
Specification
Testicle
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jul 31 2018

Application
Western blot
Sample
Guinea pig Tissue lysate - whole (Retina)
Gel Running Conditions
Reduced Denaturing (7,5%)
Loading amount
40 µg
Specification
Retina

Abcam user community

Verified customer

Submitted May 02 2018

Application
Western blot
Sample
Human Cell lysate - whole cell (human Alveolar Epithelial cells (hAELVi))
Gel Running Conditions
Reduced Denaturing (4-12%)
Loading amount
1e+006 cells
Specification
human Alveolar Epithelial cells (hAELVi)
Blocking step
Western Breeze Blocking Solution as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 0% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 30 2018

Application
Western blot
Sample
Rat Tissue lysate - whole (Liver)
Gel Running Conditions
Reduced Denaturing (10% gel)
Loading amount
10 µg
Specification
Liver
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Yu Siong Ho

Verified customer

Submitted Apr 28 2018

Application
Western blot
Sample
Mouse Cell lysate - whole cell (Testis)
Gel Running Conditions
Non-reduced Denaturing (12%)
Loading amount
40 µg
Specification
Testis
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Abcam user community

Verified customer

Submitted Jun 12 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (A549)
Gel Running Conditions
Reduced Denaturing
Loading amount
10 µg
Specification
A549
Blocking step
Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Apr 13 2017

Application
Western blot
Sample
Human Cell lysate - whole cell (B lymphoblast)
Gel Running Conditions
Reduced Denaturing (7%)
Loading amount
15 µg
Specification
B lymphoblast
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jan 20 2017

Application
Western blot
Sample
Rat Cell lysate - whole cell (Brain)
Gel Running Conditions
Reduced Denaturing (Tris-acetate 7%)
Loading amount
30 µg
Specification
Brain
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Abcam user community

Verified customer

Submitted Jun 03 2016

1-10 of 11 Abreviews or Q&A

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