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Anti-Vinculin antibody [SPM227] (ab18058)

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Overview

Product name

Anti-Vinculin antibody [SPM227]
See all Vinculin products (14) ...

Description

Mouse monoclonal [SPM227] to Vinculin

Tested applications

ICC/IF, WB, IP, IF, IHC-Pmore details

Cross reactivity

Reacts with

Mouse, Hamster, Human

Immunogen

Full length native protein (semi-purified) (Human).

Positive control

HeLa cells, skin.

Properties

Form

Liquid

Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.

Storage buffer

Preservative: 0.09% Sodium Azide
Constituents: BSA, 10mM PBS, pH 7.4

Concentration

Concentration information loading...

Purity

Protein G purified

Purification notes

Purified from ascites fluid by protein G

Clonality

Monoclonal

Clone number

SPM227

Isotype

IgG1

Light chain type

kappa

Research areas

Cardiovascular >> Heart >> Contractility >> Contractile Proteins >> Actins
Cardiovascular >> Heart >> Cardiac arrhythmias
Signal Transduction >> Cytoskeleton / ECM >> Cytoskeleton >> Microfilaments >> Actin etc >> Actin Binding Proteins

  • Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058) image (enlarge)

  • Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058) image (enlarge)

  • Western blot - Vinculin antibody [SPM227] (ab18058)Western blot - Vinculin antibody [SPM227] (ab18058) image (enlarge)

Applications

Show applications key

Our Abpromise guarantee covers the use of ab18058 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

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.

Target information above from: UniProt accessionP18206 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010).

Information by UniProt

Alternative names

  • CMD1W antibody
  • CMH15 antibody
  • Metavinculin antibody
  • MVCL antibody
  • OTTHUMP00000019861 antibody
  • OTTHUMP00000019862 antibody
  • VCL antibody
  • VINC antibody
  • VINC_HUMAN antibody
  • Vinculin antibody
see all

Database links

Anti-Vinculin antibody [SPM227] images:

  Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)

Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)

ab18058 at 1/200 staining Chinese Hamster Ovary cells by ICC/IF. The cells were permeabilized with 100% methanol and then blocked with 1% BSA/ 4% goat serum prior to incubation with the antibody for 30 minutes. A rhodamine conjugated goat antibody was used as the secondary.

This image is courtesy of an anonymous Abreview

See Abreview

  Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)

Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)

ab18058 staining mouse 3T3 cells by ICC/IF.  Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking in 1% BSA for 30 minutes at 25°C.  The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 25°C, ab6785, diluted 1/100, was used as the secondary antibody.

This image is courtesy of an Abreview submitted by Mr John Maloney

See Abreview

  Western blot - Vinculin antibody [SPM227] (ab18058)

Western blot - Vinculin antibody [SPM227] (ab18058)

Anti-Vinculin antibody [SPM227] (ab18058) at 1/1000 dilution + Whole cell lysate prepared from human lung cells at 40 µg

Secondary
HRP conjugated goat polyclonal to mouse IgG at 1/1000 dilution

Predicted band size : 130 kDa
Observed band size : 117 kDa (why is the actual band size different from the predicted?)


The image is a courtsey of an anonymous abreview.

See Abreview

  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Vinculin antibody [SPM227](ab18058)

Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Vinculin antibody [SPM227](ab18058)

Ab18058 staining human normal prostate. Staining is localised to the nucleus and cytoplasm.
Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control.
Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.

  Western blot - Vinculin antibody [SPM227] (ab18058)

Western blot - Vinculin antibody [SPM227] (ab18058)



Predicted band size : 130 kDa


Lanes: 1, 4, 7 - Hamster Lung; 2, 5, 8 - K562; 3, 6, 9 - CHO

Lanes: 1-3: 4oC (1 freeze/thaw);  4-6: 4oC (2 freeze/thaws); 7-9 :   4oC (4 freeze/thaws) 

  Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)

Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)

ICC/IF image of ab18058 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18058, 10µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

References for Anti-Vinculin antibody [SPM227] (ab18058)

This product has been referenced in:

  • Ceccaldi Ret al. Spontaneous abrogation of the G2DNA damage checkpoint has clinical benefits but promotes leukemogenesis in Fanconi anemia patients. J Clin Invest 121:184-94 (2011). WB; Human.Read more (PubMed: 21183791) »
  • Holtz ML & Misra RP Serum response factor is required for cell contact maintenance but dispensable for proliferation in visceral yolk sac endothelium. BMC Dev Biol 11:18 (2011). IHC-P; Mouse.Read more (PubMed: 21401944) »

See all 9 publications for this product

Publishing research using ab18058? Please let us know so that we can cite the reference in this datasheet

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