Vinculin Recombinant Rabbit Monoclonal Antibody [JM42-43] - BSA and Azide free
Overview
Product Name
Vinculin Recombinant Rabbit Monoclonal Antibody [JM42-43] - BSA and Azide free
Antibody Type
Recombinant Rabbit monoclonal Antibody
Immunogen
Synthetic peptide within human Vinculin aa 1,000-1,060.
Species Reactivity
Human, Mouse, Rat
Validated Applications
WB, IF-Cell, IF-Tissue, IHC-P
Molecular Weight
Predicted band size: 124 kDa
Positive Control
HepG2 cell lysate, HeLa cell lysate, NIH/3T3 cell lysate, C6 cell lysate, Mouse spleen tissue lysate, Rat kidney tissue lysate, NIH/3T3, C6, HeLa, mouse spleen tissue, mouse testis tissue, rat colon tissue, rat spleen tissue, rat testis tissue.
Conjugation
unconjugated
Clone Number
JM42-43
Product Features
Form
Liquid
Concentration
Storage Instructions
Store at +4℃ after thawing. Aliquot store at -20℃ or -80℃. Avoid repeated freeze / thaw cycles.
Storage Buffer
1*PBS (pH7.4).
Isotype
IgG
Purification Method
Protein A affinity purified.
Application Dilution
-
WB
-
1:20,000-1:50,000
-
IF-Cell
-
1:200-1:1,000
-
IF-Tissue
-
1:500-1:4,000
-
IHC-P
-
1:5,000-1:20,000
Target
Function
Focal adhesions are identified as areas within the plasma membrane of tissue culture cells that adhere tightly to the underlying substrate. In vivo, these regions are involved in the adhesion of cells to the extracellular matrix. Paxillin and vinculin are cytoskeletal, focal adhesion proteins that are components of a protein complex which links the Actin network to the plasma membrane. Vinculin binding sites have been identified on other cytoskeletal proteins, including Talin and α-actinin. In addition, vinculin, Talin and α-actinin each contain Actin binding sites. Expression of vinculin and Talin have been shown to be affected by the level of Actin expression. α-Actinin has been shown to link Actin to integrins in the plasma membrane through interactions with the vinculin and Talin complex or by a direct interaction with integrin.
Background References
1. King HO et al. RAD51 Is a Selective DNA Repair Target to Radiosensitize Glioma Stem Cells. Stem Cell Reports 8:125-139 (2017).
2. Boyette LB et al. Phenotype, function, and differentiation potential of human monocyte subsets. PLoS One 12:e0176460 (2017).
Sequence Similarity
Belongs to the vinculin/alpha-catenin family.
Tissue Specificity
Metavinculin is muscle-specific.
Post-translational Modification
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 (By similarity).; Acetylated; mainly by myristic acid but also by a small amount of palmitic acid.
Subcellular Location
Cell junction, Cell membrane, Cytoplasm, Cytoskeleton, Membrane.
Synonyms
CMD1W antibody
CMH15 antibody
Epididymis luminal protein 114 antibody
HEL114 antibody
Metavinculin antibody
MV antibody
MVCL antibody
OTTHUMP00000019861 antibody
OTTHUMP00000019862 antibody
VCL antibody
ExpandImages
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Western blot analysis of Vinculin on different lysates with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
Lane 1: HepG2 cell lysate (15 µg/Lane)
Lane 2: HeLa cell lysate (15 µg/Lane)
Lane 3: NIH/3T3 cell lysate (15 µg/Lane)
Lane 4: C6 cell lysate (15 µg/Lane)
Lane 5: Mouse spleen tissue lysate (20 µg/Lane)
Lane 6: Rat kidney tissue lysate (20 µg/Lane)
Predicted band size: 124 kDa
Observed band size: 124 kDa
Exposure time: 1 minute 21 seconds;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750448) at 1/20,000 dilution was used in 5% NFDM/TBST at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
☑ Knockdown (KD)
Western blot analysis of Vinculin on different lysates with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
Lane 1: HAP1-parental cell lysate
Lane 2: HAP1-Vinculin KD cell lysate
Lysates/proteins at 10 µg/Lane.
Predicted band size: 124 kDa
Observed band size: 124 kDa
Exposure time: 10 seconds; ECL: K1801;
4-20% SDS-PAGE gel.
Proteins were transferred to a PVDF membrane and blocked with 5% NFDM/TBST for 1 hour at room temperature. The primary antibody (HA750448) at 1/20,000 dilution was used in K1803 at 4℃ overnight. Goat Anti-Rabbit IgG - HRP Secondary Antibody (HA1001) at 1/50,000 dilution was used for 1 hour at room temperature. -
Immunocytochemistry analysis of NIH/3T3 cells labeling Vinculin with Rabbit anti-Vinculin antibody (HA750448) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Vinculin antibody (HA750448) at 1/1250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of C6 cells labeling Vinculin with Rabbit anti-Vinculin antibody (HA750448) at 1/250 dilution.
Cells were fixed in 4% paraformaldehyde for 20 minutes at room temperature, permeabilized with 0.1% Triton X-100 in PBS for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Vinculin antibody (HA750448) at 1/1250 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI.
Beta tubulin (M1305-2, red) was stained at 1/100 dilution overnight at +4℃. Goat Anti-Mouse IgG H&L (iFluor™ 594, HA1126) was used as the secondary antibody at 1/1,000 dilution. -
Immunocytochemistry analysis of HeLa cells labeling Vinculin with Rabbit anti-Vinculin antibody (HA750448) at 1/1,000 dilution.
Cells were fixed in 100% precooled methanol for 5 minutes at room temperature, then blocked with 1% BSA in 10% negative goat serum for 1 hour at room temperature. Cells were then incubated with Rabbit anti-Vinculin antibody (HA750448) at 1/1,000 dilution in 1% BSA in PBST overnight at 4 ℃. Goat Anti-Rabbit IgG H&L (iFluor™ 488, HA1121) 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. Nuclear DNA was labelled in blue with DAPI. -
Immunohistochemical analysis of paraffin-embedded mouse spleen tissue with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750448) at 1/20,000 dilution for 1 hour at room temperature. 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 with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750448) at 1/20,000 dilution for 1 hour at room temperature. 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 rat colon tissue with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750448) at 1/20,000 dilution for 1 hour at room temperature. 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 rat spleen tissue with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750448) at 1/20,000 dilution for 1 hour at room temperature. 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 rat testis tissue with Rabbit anti-Vinculin antibody (HA750448) at 1/20,000 dilution.
The section was pre-treated using heat mediated antigen retrieval with Tris-EDTA buffer (pH 9.0) for 20 minutes. The tissues were blocked in 1% BSA for 20 minutes at room temperature, washed with ddH2O and PBS, and then probed with the primary antibody (HA750448) at 1/20,000 dilution for 1 hour at room temperature. 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. -
Flow cytometric analysis of HeLa cells labeling Vinculin.
Cells were fixed and permeabilized. Then stained with the primary antibody (HA750448, 1μg/mL) (red) compared with Rabbit IgG Isotype Control (green). After incubation of the primary antibody at +4℃ for an hour, the cells were stained with a iFluor™ 488 conjugate-Goat anti-Rabbit IgG Secondary antibody (HA1121) at 1/1,000 dilution for 30 minutes at +4℃. Unlabelled sample was used as a control (cells without incubation with primary antibody; black).
Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"
Citation
-
A Novel CSN5 Inhibitor Drives Tumor-Intrinsic PD-L1 Degradation and Exerts Direct Antitumor Efficacy in Triple-Negative Breast Cancer
Journal: Journal Of Medicinal Chemistry
DOI: 10.1021/acs.jmedchem.5c02669
IF: 6.8
Application: WB
Reactivity: Human
Publish date: 2026 Feb
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