Vinculin Antibody (YA010)

Vinculin Antibody is a non-conjugated and Rabbit origined IgG monoclonal antibody, targeting to Vinculin. It can be used as a loading control antibody.

Vinculin, a crucial actin filament (F-actin)-binding protein, actively participates in cell-matrix adhesion and cell-cell adhesion processes. It plays a regulatory role in the expression of cell-surface E-cadherin and enhances mechanosensing by the E-cadherin complex, contributing to cell morphology and locomotion (By similarity). Notably, vinculin exhibits self-association properties and is part of a complex that includes THSD1, PTK2/FAK1, TLN1, and VCL (By similarity). Its interactions extend to various proteins such as APBB1IP, NRAP, TLN1, SYNM, CTNNB1, SORBS1, and CTNNA1, each interaction serving specific cellular functions. The interaction with CTNNB1 is particularly essential for vinculin's localization to cell-cell junctions and its role in regulating the cell surface expression of E-cadherin (By similarity). Additionally, vinculin binds to ACTN4, triggering conformational changes that further contribute to its functional versatility (By similarity).

Free

P18206

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

Protein A affinity purified.

Cell junction, Cell membrane, Cytoplasm, Cytoskeleton, Membrane.

Non-conjugated

Unmodified

AB_3102462

Signal Transduction

Primary Antibody; Recombinant Rabbit Monoclonal Antibody

Recombinant, Monoclonal

Rabbit

Human, Mouse, Rat

WB: 1:500-1:2,000 ;ICC/IF: 1:50-1:200 ;IP: Use at an assay dependent concentration. ;IHC: 1:200

• 
  Western blot analysis of extracts from HepG2 (lane 2(20μg) , Hela(lane 3(20μg) and NIH3T3(lane 4(20μg) using Vinculin(HY-P80372) Rabbit mAb. Proteins were transferred to a PVDF membrane and blocked with 5% non-fat milk in TBST for 2 hour at room temperature. The primary antibody (1/1000) and Loading control antibody (Beta Actin, HY-P80438, 1/10000) was used in 5% non-fat milk in TBST at 4°C overnight. Goat Anti-Mouse IgG-HRP Secondary Antibody (1/10000) was used for 1 hour at room temperature.
• 
  Immunocytochemistry analysis of HUVEC cells labeling Vinculin with Vinculin Antibody (HY-P80372)at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Vinculin Antibody (HY-P80372) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunocytochemistry analysis of Hela cells labeling Vinculin with Vinculin Antibody (HY-P80372) at 1/50 dilution. Cells were fixed in 4% paraformaldehyde for 15 minutes at room temperature, permeabilized with 0.1% Triton X-100 for 10 minutes at room temperature, then blocked with QuickBlock™ Blocking Buffer for Immunol Staining for 10 min at room temperature. Cells were then incubated with Vinculin Antibody (HY-P80372) at 1/50 dilution in QuickBlock™ Blocking Buffer for Immunol Staining at 4 ℃. Alexa Fluor® 488-conjugated AffiniPure Goat Anti-Rabbit IgG H&L（HY-P8002, Green) was used as the secondary antibody at 1/1,000 dilution. PBS instead of the primary antibody was used as the secondary antibody only control. The Nuclear counterstain was DAPI (Blue).
• 
  Immunohistochemical analysis of paraffin-embedded Mouse testis tissue using Vinculin Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80372, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.
• 
  Immunohistochemical analysis of paraffin-embedded Mouse testis tissue using Vinculin Antibody. The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 8 minutes. The tissues were blocked in QuickBlock for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HY-P80372, 1/1000) in 4℃ overnight. The detection was performed using an HRP conjugated compact polymer system. DAB was used as the chromogen. Tissues were counterstained with hematoxylin and mounted with DPX.

WB, IHC-P, ICC/IF

Liquid

Supplied in 1*TBS (pH7.4), 0.05% BSA and 40% Glycerol. Preservative: 0.05% Sodium Azide.

Stored at -20°C for 1 year. Avoid repeated freeze / thaw cycles.

Shipping with blue ice.

IgG

Endogenous

Synthetic peptide corresponding to Human Vinculin.AA range:1000-1060.