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    products/primary-antibodies/vinculin-antibody-epr8185-ab129002.pdf

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RecombinantRabMAb

Recombinant Anti-Vinculin antibody [EPR8185] (ab129002)

  • Datasheet
  • SDS
Reviews (19) Submit a question References (206)

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Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Flow Cytometry (Intracellular) - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Immunoprecipitation - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
  • Anti-Vinculin antibody [EPR8185] (ab129002)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR8185] to Vinculin
  • Suitable for: Flow Cyt (Intra), WB, IP, ICC/IF
  • Reacts with: Mouse, Rat, Human

Conjugates logo Related conjugates and formulations

Alexa Fluor® 488 Alexa Fluor® 555 Alexa Fluor® 647 Carrier Free

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View more associated products

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 (Intra), WB, IP, ICC/IFmore details
    Unsuitable for: IHC-P
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • 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 (intra): HEK293 cells.
  • 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.

    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.

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Stable for 12 months at -20°C.
  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 40% Glycerol, 0.05% BSA, 59% PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EPR8185
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Binding Proteins
    • Cardiovascular
    • Heart
    • Cardiac arrhythmias
    • Cardiovascular
    • Heart
    • Contractility
    • Contractile Proteins
    • Actins
    • Isotype/Loading Controls
    • Loading Controls
    • Vinculin

Associated products

  • Alternative Versions

    • Alexa Fluor® 488 Anti-Vinculin antibody [EPR8185] (ab196454)
    • Alexa Fluor® 647 Anti-Vinculin antibody [EPR8185] (ab196579)
    • Alexa Fluor® 555 Anti-Vinculin antibody [EPR8185] - Loading Control (ab206908)
    • Anti-Vinculin antibody [EPR8185] - BSA and Azide free (ab217171)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • Positive Controls

    • U937 cytoplasmic extract lysate (ab14904)
  • Recombinant Protein

    • Recombinant Human Vinculin protein (ab131747)
  • Related Products

    • Prestained Protein Ladder – Broad molecular weight (10-245 kDa) (ab116028)

Applications

The Abpromise guarantee

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 (Intra)
1/200.
WB (16)
1/10000 - 1/50000. Detects a band of approximately 124 kDa (predicted molecular weight: 124 kDa).
IP
1/10 - 1/100.
ICC/IF (2)
1/50 - 1/250.
Notes
Flow Cyt (Intra)
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.
Application notes
Is unsuitable for IHC-P.

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 accession P18206 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7414 Human
    • Entrez Gene: 22330 Mouse
    • Entrez Gene: 305679 Rat
    • Omim: 193065 Human
    • SwissProt: P18206 Human
    • SwissProt: Q64727 Mouse
    • SwissProt: P85972 Rat
    • Unigene: 643896 Human
    • Unigene: 279361 Mouse
    • Unigene: 164613 Rat
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] (ab129002)
    Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] (ab129002)

    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.

  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    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 (Intracellular) - Anti-Vinculin antibody [EPR8185] (ab129002)
    Flow Cytometry (Intracellular) - Anti-Vinculin antibody [EPR8185] (ab129002)

    Intracellular 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 Fluorr® 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. 

     

     

  • Immunoprecipitation - Anti-Vinculin antibody [EPR8185] (ab129002)
    Immunoprecipitation - Anti-Vinculin antibody [EPR8185] (ab129002)

    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.

  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    Different batches of ab129002 were tested on HepG2 (Human hepatocellular carcinoma epithelial cell) lysate at 1.0 µg/ml. 15 µg of lysate was loaded in each lane. Bands observed at 124 kDa.
  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    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

  • Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] (ab129002)
    Immunocytochemistry/ Immunofluorescence - Anti-Vinculin antibody [EPR8185] (ab129002)

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

  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    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

  • Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    Western blot - Anti-Vinculin antibody [EPR8185] (ab129002)
    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

  • Anti-Vinculin antibody [EPR8185] (ab129002)
    Anti-Vinculin antibody [EPR8185] (ab129002)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (206)

Publishing research using ab129002? Please let us know so that we can cite the reference in this datasheet.

ab129002 has been referenced in 206 publications.

  • Caputo T  et al. Anti-adipogenic signals at the onset of obesity-related inflammation in white adipose tissue. Cell Mol Life Sci 78:227-247 (2021). PubMed: 32157317
  • Moretti CH  et al. Germ-free mice are not protected against diet-induced obesity and metabolic dysfunction. Acta Physiol (Oxf) 231:e13581 (2021). PubMed: 33222397
  • Hervás-Salcedo R  et al. Enhanced anti-inflammatory effects of mesenchymal stromal cells mediated by the transient ectopic expression of CXCR4 and IL10. Stem Cell Res Ther 12:124 (2021). PubMed: 33579367
  • Baranska E  et al. Interactions of a Water-Soluble Glycofullerene with Glucose Transporter 1. Analysis of the Cellular Effects on a Pancreatic Tumor Model. Nanomaterials (Basel) 11:N/A (2021). PubMed: 33670509
  • Sannomiya Y  et al. The role of discoidin domain receptor 2 in the renal dysfunction of alport syndrome mouse model. Ren Fail 43:510-519 (2021). PubMed: 33706638
View all Publications for this product

Customer reviews and Q&As

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1-10 of 19 Abreviews or Q&A

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (Fibroblasts)
Gel Running Conditions
Reduced Denaturing
Loading amount
15 µg
Specification
Fibroblasts
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 29 2021

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Zebrafish Tissue lysate - whole (whole embryo)
Gel Running Conditions
Reduced Denaturing (4–20% precast polyacrylamide gel))
Loading amount
40 µg
Specification
whole embryo
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Nov 02 2021

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (pancreatic cancer)
Gel Running Conditions
Reduced Denaturing (BioRad MiniProtean precast gels)
Loading amount
40 µg
Specification
pancreatic cancer
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

DR. Soledad Camolotto

Verified customer

Submitted Apr 01 2021

Immunohistochemistry (PFA perfusion fixed frozen sections) abreview for Anti-Vinculin antibody [EPR8185]

Good
Abreviews
Abreviews
abreview image
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Mouse Tissue sections (Brain (Cortex))
Antigen retrieval step
None
Permeabilization
Yes - PBS 1x, Triton x-100 1%
Specification
Brain (Cortex)
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Paraformaldehyde
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Dec 08 2020

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Mouse Tissue lysate - whole (Cerebellum)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
40 µg
Specification
Cerebellum
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 27 2020

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Rat Cell lysate - whole cell (Hippocampal culture)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
40 µg
Specification
Hippocampal culture
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Feb 27 2020

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
Application
Western blot
Sample
Human Cell lysate - whole cell (HEK293)
Gel Running Conditions
Reduced Denaturing (4-15% gradient)
Loading amount
20 µg
Specification
HEK293
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Feb 27 2020

Immunocytochemistry/ Immunofluorescence abreview for Anti-Vinculin antibody [EPR8185]

Good
Abreviews
Abreviews
abreview image
Application
Immunocytochemistry/ Immunofluorescence
Sample
Pig Cell (Kidney)
Permeabilization
Yes - Heat fixation
Specification
Kidney
Fixative
Heat fixation
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Verified customer

Submitted Feb 27 2020

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

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Submitted Jan 08 2019

Immunocytochemistry/ Immunofluorescence abreview for Anti-Vinculin antibody [EPR8185]

Excellent
Abreviews
Abreviews
abreview image
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
Read More
The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Anugraha Rajagopalan

Verified customer

Submitted Aug 17 2018

1-10 of 19 Abreviews or Q&A

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