Name: Anti-Vinculin antibody [EPR8185]
Brand: RabMAb
SKU: ab129002
Rating: 4 (19 reviews)

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Immunocytochemistry/ Immunofluorescence - 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

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

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.

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

Alternative Versions
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)

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.

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

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)

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)

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)

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)

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)

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)

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

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)

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

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

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

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

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

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

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

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

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

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

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 Dec 08 2020

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

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

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

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

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

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

Application

Immunocytochemistry/ Immunofluorescence

Sample

Pig Cell (Kidney)

Permeabilization

Yes - Heat fixation

Specification

Kidney

Fixative

Heat fixation

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

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

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 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

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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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Positive Controls

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

References (206)

Customer reviews and Q&As

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Western blot abreview for Anti-Vinculin antibody [EPR8185]

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

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Western blot abreview for Anti-Vinculin antibody [EPR8185]

Western blot abreview for Anti-Vinculin antibody [EPR8185]

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

Western blot abreview for Anti-Vinculin antibody [EPR8185]

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

Post-translational
modifications