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Purified anti-Vinculin Antibody

Pricing & Availability
Clone
W18245A (See other available formats)
Regulatory Status
RUO
Other Names
Metavinculin, MV, Epididymis Secretory Sperm Binding Protein, Epididymis Luminal Protein 114
Isotype
Rat IgG2a, κ
Ave. Rating
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Product Citations
publications
1. W18245A_PURE_Vinculin_Antibody_1_072720
Whole cell extracts (15 µg protein) from PC-3, HeLa, MOLT-4 (reduced expression control), and NIH/3T3 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-Vinculin antibody (clone W18245A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker. Predicted VCL expression data was obtained from Human Protein Atlas.
  • 1. W18245A_PURE_Vinculin_Antibody_1_072720
    Whole cell extracts (15 µg protein) from PC-3, HeLa, MOLT-4 (reduced expression control), and NIH/3T3 cells were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with 1.0 µg/mL (1:500 dilution) of purified anti-Vinculin antibody (clone W18245A) overnight at 4°C. Proteins were visualized by chemiluminescence detection using HRP goat anti-rat IgG antibody (Cat. No. 405405) at a 1:3000 dilution. Direct-Blot™ HRP anti-GAPDH antibody (Cat. No. 607904) was used as a loading control at a 1:25000 dilution (lower). Lane M: Molecular weight marker. Predicted VCL expression data was obtained from Human Protein Atlas.
  • 2. W18245A_PURE_Vinculin_Antibody_2_072720
    HeLa cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with ice-cold methanol for 10 minutes, and blocked with 5% FBS for 60 minutes. Cells were then intracellularly stained with 5.0 µg/mL (1:100 dilution) of either purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400501) (panel A) or purified anti-Vinculin antibody (panel B) for two hours at room temperature, followed by incubation with Alexa Fluor™ 594 goat anti-rat IgG (Cat. No. 405422) at 2.0 µg/mL. Nuclei were counterstained with DAPI, and the image was captured with a 60X objective.
  • 3. W18245A_PURE_Vinculin_Antibody_3_072720
    Whole cell extracts (250 µg total protein) prepared from HeLa cells were immunoprecipitated overnight with 2.0 µg of purified rat IgG2a, κ isotype ctrl antibody (Cat. No. 400501) or purified anti-Vinculin antibody (clone W18245A). The resulting IP fractions and whole cell extract input (6%) were resolved by 4-12% Bis-Tris gel electrophoresis, transferred to a PVDF membrane, and probed with a rabbit control antibody raised against a separate epitope of Vinculin. Lane M: Molecular weight marker.
  • 4. W18245A_PURE_Vinculin_Antibody_072722
    IHC staining using purified anti-Vinculin (clone W18245A) on formalin-fixed paraffin-embedded human heart tissue. Following antigen retrieval using 1X Tris-Buffered Saline (final concentration 0.05M) with Tween-20 (Cat. No. 925501), the tissue was incubated with (B) or without (A) 10 μg/mL of antibody overnight at 4°C, followed by incubation with 2.5 μg/mL of Alexa Fluor® 647 goat anti-rat IgG (Cat. No. 405416) for one hour at room temperature. Nuclei were counterstained with DAPI (blue) (Cat. No. 422801), and the slide was mounted with ProLong™ Gold Antifade Mountant. The image was captured with a 40X objective. Scalebar = 50 μM
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938401 25 µg $101
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938402 100 µg $253
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Description

Vinculin is a filamentous actin (F-actin) binding protein that was originally described as a critical component of focal adhesion plaques. It plays an important role in the transmission of mechanical forces, stabilization of cell-cell junctions via a direct interaction with α-catenin, and linking of integrins to the actin cytoskeleton. Loss of Vinculin protein expression contributes to metastatic phenotypes in cancer tissue due to loss of cell-cell adhesion. Vinculin has been isolated from prostate cancer exosomes and may serve as a biomarker for the progression of taxane-resistance prostate cancers; it is also a common marker for cell-cell and focal adhesion junctions.

Product Details
Technical data sheet

Product Details

Verified Reactivity
Human, Mouse
Antibody Type
Monoclonal
Host Species
Rat
Immunogen
Partial recombinant human Vinculin protein
Formulation
Phosphate-buffered solution, pH 7.2, containing 0.09% sodium azide
Preparation
The antibody was purified by affinity chromatography.
Concentration
0.5 mg/mL
Storage & Handling
The antibody solution should be stored undiluted between 2°C and 8°C.
Application

WB - Quality tested
ICC, IP, IHC-P - Verified
Recommended Usage

Each lot of this antibody is quality control tested by western blotting. For western blotting, the suggested use of this reagent is 0.1 - 1.0 µg/mL. For immunocytochemistry, a concentration range of 1.0 - 5.0 μg/mL is recommended. For immunoprecipitation, the suggested use of this reagent is 2.0 µg/test. For immunohistochemistry on formalin-fixed paraffin-embedded tissue sections, a concentration range of 5.0 - 10.0 µg/mL is suggested. It is recommended that the reagent be titrated for optimal performance for each application.

Application Notes

This clone was tested for ICC using HeLa cells fixed with 4% PFA and permeabilized with Triton X-100 or methanol. While methanol permeabilization was compatible with Vinculin staining, Triton X-100 permeabilization resulted in dim staining. We therefore do not recommend Triton X-100 as a permeabilization method.

While this clone passed IP testing, it failed to quantitatively enrich Vinculin from whole cell extracts and may not be suitable for detecting transient or low-abundance protein-protein interactions by co-immunoprecipitation.

This clone is predicted to recognize Metavinculin due to complete sequence homology between the immunogen and the corresponding region of Metavinculin.

RRID
AB_2876761 (BioLegend Cat. No. 938401)
AB_2876761 (BioLegend Cat. No. 938402)

Antigen Details

Structure
Vinculin is a 1,066 amino acid protein with a predicted molecular weight of 116 kD. Metavinculin is a 1,134 amino acid protein with a predicted molecular weight of 124 kD.
Distribution

Ubiquitously expressed/cytosol, cytoskeleton, and focal adhesion plaques; Metavinculin expression is restricted to smooth skeletal muscle tissue.

Function
Cell-matrix and cell-cell adhesion
Biology Area
Cell Biology, Cell Motility/Cytoskeleton/Structure
Molecular Family
Cytoskeletal Proteins
Antigen References
  1. Kawakami K, et al. 2015. Int J Oncol. 47:384-90.
  2. Geiger B, et al. 1980. Proc Natl Acad Sci U S A. 77:4127-31.
  3. Jockusch BM, et al. 1981. Proc Natl Acad Sci U S A. 78:3005-9.
Gene ID
7414 View all products for this Gene ID
UniProt
View information about Vinculin on UniProt.org

Related FAQs

There are no FAQs for this product.
Go To Top Version: 2    Revision Date: 07/27/2022

For Research Use Only. Not for diagnostic or therapeutic use.

 

This product is supplied subject to the terms and conditions, including the limited license, located at www.biolegend.com/terms) ("Terms") and may be used only as provided in the Terms. Without limiting the foregoing, BioLegend products may not be used for any Commercial Purpose as defined in the Terms, resold in any form, used in manufacturing, or reverse engineered, sequenced, or otherwise studied or used to learn its design or composition without express written approval of BioLegend. Regardless of the information given in this document, user is solely responsible for determining any license requirements necessary for user’s intended use and assumes all risk and liability arising from use of the product. BioLegend is not responsible for patent infringement or any other risks or liabilities whatsoever resulting from the use of its products.

 

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This data display is provided for general comparisons between formats.
Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors.
If you need assistance with selecting the best format contact our expert technical support team.

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