Recombinant Anti-Vinculin antibody [EPR20407] KO Tested (ab219649)

Product name

Anti-Vinculin antibody [EPR20407]
See all Vinculin primary antibodies
Description

Rabbit monoclonal [EPR20407] to Vinculin
Host species

Rabbit
Tested applications

Suitable for: WB, IHC-Pmore details
Species reactivity

Reacts with: Mouse, Rat, Human
Immunogen

Recombinant fragment. This information is proprietary to Abcam and/or its suppliers.
Positive control
- WB: PC-3, HeLa, K562, C6, A431, and PC-12 whole cell lysates; Human fetal kidney, fetal heart, testis and fetal liver lysates; Mouse heart and spleen lysates; Rat heart, kidney and spleen lysates. IHC-P: Human breast, colon, gastric cancer and prostate cancer tissues; Mouse colon tissue; Rat stomach tissue.
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.

Form

Liquid
Storage instructions

Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
Storage buffer

pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20407
Isotype

IgG
Research areas

Alternative Versions
- Anti-Vinculin antibody [EPR20407] - BSA and Azide free (ab240395)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
KO cell lines
- Human VCL (Vinculin) knockout A-431 cell line (ab261898)
KO cell lysates
- Human VCL (Vinculin) knockout A-431 cell lysate (ab261707)
Recombinant Protein
- Recombinant Human Vinculin protein (ab131747)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab219649 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		1/1000. Detects a band of approximately 123 kDa (predicted molecular weight: 124 kDa).
IHC-P		1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

Notes
WB 1/1000. Detects a band of approximately 123 kDa (predicted molecular weight: 124 kDa).
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

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

Western blot - Anti-Vinculin antibody [EPR20407] (ab219649)

All lanes : Anti-Vinculin antibody [EPR20407] (ab219649) at 1/1000 dilution

Lane 1 : Wild-type A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 2 : VCL knockout A-431 (Human epidermoid carcinoma cell line) whole cell lysate
Lane 3 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 4 : Jurkat (Human T cell leukemia cell line from peripheral blood) whole cell lysate

Lysates/proteins at 20 µg per lane.

Performed under reducing conditions.

Predicted band size: 124 kDa
Observed band size: 124 kDa

Lanes 1 - 4: Merged signal (red and green). Green - ab219649 observed at 124 kDa. Red - loading control, ab8245 (Mouse anti-GAPDH antibody [6C5]) observed at 37kDa.

ab219649 was shown to react with VCL in A431 wild-type cells in Western blot. Loss of signal was observed when VCL knockout sample was used. A431 wild-type and VCL knockout cell lysates were subjected to SDS-PAGE. Membranes were blocked in 3% Milk in TBS-T (0.1% Tween®) before incubation with ab219649 and ab8245 (Mouse anti-GAPDH antibody [6C5]) overnight at 4°C at a 1 in 1000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vinculin antibody [EPR20407] (ab219649)

Immunohistochemical analysis of paraffin-embedded human breast tissue labeling Vinculin with ab219649 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous staining on myoepithelium of human breast is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vinculin antibody [EPR20407] (ab219649)

Immunohistochemical analysis of paraffin-embedded human colon tissue labeling Vinculin with ab219649 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous and cytoplasmic staining on smooth muscle cells of human colon is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Western blot - Anti-Vinculin antibody [EPR20407] (ab219649)

All lanes : Anti-Vinculin antibody [EPR20407] (ab219649) at 1/1000 dilution

Lane 1 : PC-3 (Human prostate adenocarcinoma cell line) whole cell lysate
Lane 2 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
Lane 3 : K562 (Human chronic myelogenous leukemia cell line from bone marrow) whole cell lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 124 kDa
Observed band size: 123 kDa why is the actual band size different from the predicted?

Exposure time: 15 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Western blot - Anti-Vinculin antibody [EPR20407] (ab219649)

All lanes : Anti-Vinculin antibody [EPR20407] (ab219649) at 1/1000 dilution

Lane 1 : Human fetal kidney lysate
Lane 2 : Human fetal heart lysate
Lane 3 : Human testis lysate
Lane 4 : Human fetal liver lysate

Lysates/proteins at 20 µg per lane.

Secondary
All lanes : VeriBlot for IP Detection Reagent (HRP) (ab131366) at 1/4000 dilution

Predicted band size: 124 kDa
Observed band size: 123 kDa why is the actual band size different from the predicted?

Blocking/Dilution buffer: 5% NFDM/TBST.

Exposure time: Lane 1-3: 15 seconds; Lane 4: 30 seconds.
Western blot - Anti-Vinculin antibody [EPR20407] (ab219649)

All lanes : Anti-Vinculin antibody [EPR20407] (ab219649) at 1/1000 dilution

Lane 1 : Mouse heart lysate
Lane 2 : Mouse spleen lysate
Lane 3 : Rat heart lysate
Lane 4 : Rat kidney lysate
Lane 5 : Rat spleen lysate
Lane 6 : C6 (Rat glial tumor cell line) whole cell lysate
Lane 7 : PC-12 (Rat adrenal gland pheochromocytoma cell line) whole cell lysate

Lysates/proteins at 10 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

Predicted band size: 124 kDa
Observed band size: 123 kDa why is the actual band size different from the predicted?

Exposure time: 4 seconds

Blocking/Dilution buffer: 5% NFDM/TBST.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vinculin antibody [EPR20407] (ab219649)

Immunohistochemical analysis of paraffin-embedded human gastric cancer tissue labeling Vinculin with ab219649 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous and cytoplasmic staining on smooth muscle cells of human gastric cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vinculin antibody [EPR20407] (ab219649)

Immunohistochemical analysis of paraffin-embedded human prostate cancer tissue labeling Vinculin with ab219649 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous and cytoplasmic staining on smooth muscle cells of human prostate cancer is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vinculin antibody [EPR20407] (ab219649)

Immunohistochemical analysis of paraffin-embedded mouse colon tissue labeling Vinculin with ab219649 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous and cytoplasmic staining on smooth muscle cells of mouse colon is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vinculin antibody [EPR20407] (ab219649)

Immunohistochemical analysis of paraffin-embedded rat stomach tissue labeling Vinculin with ab219649 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Membranous and cytoplasmic staining on smooth muscle cells of rat stomach is observed.

Counter stained with Hematoxylin.

Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Anti-Vinculin antibody [EPR20407] (ab219649)

Western blot protocols
Immunohistochemistry protocols

Click here to view the general protocols

SDS download

Country/Region

Language
Datasheet download

Download

Publishing research using ab219649? Please let us know so that we can cite the reference in this datasheet.

ab219649 has been referenced in 7 publications.

Lappin KM et al. Cancer-Associated SF3B1 Mutations Confer a BRCA-Like Cellular Phenotype and Synthetic Lethality to PARP Inhibitors. Cancer Res 82:819-830 (2022). PubMed: 35027467
Zhou J et al. The Expression of Cytokine Profiles and Related Receptors in Idiopathic Inflammatory Myopathies. Front Pharmacol 13:852055 (2022). PubMed: 35517781
Jin H et al. A Fluorescence-Microscopic System for Monitoring the Turnover of the Autophagic Substrate p62/SQSTM1. Methods Mol Biol 2543:71-82 (2022). PubMed: 36087260
Milighetti M et al. Proteomic profiling of soft tissue sarcomas with SWATH mass spectrometry. J Proteomics 241:104236 (2021). PubMed: 33895336
Nouws J et al. MicroRNA miR-24-3p reduces DNA damage responses, apoptosis, and susceptibility to chronic obstructive pulmonary disease. JCI Insight 6:N/A (2021). PubMed: 33290275
Sheng J et al. Identification by Comprehensive Bioinformatics Analysis of KIF15 as a Candidate Risk Gene for Triple-Negative Breast Cancer. Cancer Manag Res 12:12337-12348 (2020). PubMed: 33293861
Zhang M et al. Vinculin promotes gastric cancer proliferation and migration and predicts poor prognosis in patients with gastric cancer. J Cell Biochem 120:14107-14115 (2019). PubMed: 30989694

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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

KO cell lines

KO cell lysates

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

Certificate of Compliance

References (7)

Customer reviews and Q&As

Post-translational
modifications