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Immunofluorescence analysis of Vinculin was performed using 70% confluent log phase U-2 OS cells. The cells were fixed with 4% paraformaldehyde for 10 minutes, permeabilized with 0.1% Triton™ X-100 for 10 minutes, and blocked with 1% BSA for 1 hour at room temperature. The cells were labeled with Vinculin (VLN01) Mouse Monoclonal Antibody (MA5-11690) at 2 µg/ml in 0.1% BSA and incubated for 3 hours at room temperature and then labeled with Goat anti-Mouse IgG (H+L) Superclonal™ Secondary Antibody, Alexa Fluor® 488 conj µgate (A28175) at a dilution of 1:2000 for 45 minutes at room temperature (Panel a: green). Nuclei (Panel b: blue) were stained with SlowFade® Gold Antifade Mountant with DAPI (S36938). F-actin (Panel c: red) was stained with Alexa Fluor® 555 Rhodamine Phalloidin (Product # R415, 1:300). Panel d represents the merged image showing cytoplasmic localization. Panel e shows the no primary antibody control. The images were captured at 60X magnification.
|Tested species reactivity||Human, Mouse, Primate|
|Published species reactivity||Cat, Human|
|Host / Isotype||Mouse / IgG|
|Storage buffer||PBS, pH 7.4, with 0.2% BSA|
|Contains||0.09% sodium azide|
|Storage Conditions||4° C|
|Tested Applications||Dilution *|
|Flow Cytometry (Flow)||3-5 µg/10^6 cells|
|Immunocytochemistry (ICC)||2 µg/ml|
|Immunofluorescence (IF)||2 µg/mL|
|Immunohistochemistry (Paraffin) (IHC (P))||1:50|
|Immunoprecipitation (IP)||2 µg/ml|
|Western Blot (WB)||1-3 µg/mL|
* Suggested working dilutions are given as a guide only. It is recommended that the user titrate the product for use in their own experiment using appropriate negative and positive controls.
MA5-11690 targets Vinculin in IF, IHC (P), IP, and WB applications and shows reactivity with Human samples.
The MA5-11690 immunogen is semi-purified vinculin.
Vinculin is an attachment protein involved in the indirect binding of intracellular actin filaments to extracellular fibronectin. Vinculin binds to actin capping protein and alpha actinin, which in turn binds to actin. The binding of fibronectin is also indirect where vinculin binds to talin, which in turn binds to the intracellular domain of the fibronectin receptor which itself is the ligand for fibronectin. Vinculin is widely distributed in tissue and is expressed where smooth muscle actin and fibroblasts attach to the extracellular matrix.
For Research Use Only. Not for use in diagnostic procedures. Not for resale without express authorization.
Focal adhesion plaque associated cytoskeletons are involved in the invasion and metastasis of human colorectal carcinoma.
MA5-11690 was used in immunohistochemistry to investigate the involvement of focal adhesion plaque associated cytoskeletons in the invasion and metastasis of colorectal carcinoma
|Yang HJ,Chen JZ,Zhang WL,Ding YQ||Cancer investigation (28:127)||2010|
Connective tissue growth factor expression and induction by transforming growth factor-beta is abrogated by simvastatin via a Rho signaling mechanism.
MA5-11690 was used in western blot to investigate the influence of simvastatin on connective tissue growth factor expression and its mechanism
|Watts KL,Spiteri MA||American journal of physiology. Lung cellular and molecular physiology (287:L1323)||2004|
Focal complex formation in adult cardiomyocytes is accompanied by the activation of beta3 integrin and c-Src.
MA5-11690 was used in immunocytochemistry to investigate the involvement of beta3 integrin and c-Src in focal adhesion complex formation in adult cardiomyocytes
|Willey CD,Balasubramanian S,Rodríguez Rosas MC,Ross RS,Kuppuswamy D||Journal of molecular and cellular cardiology (35:671)||2003|
epididymis luminal protein 114; metavinculin; vinculin
9430097D22; AA571387; AI462105; AW545629; CMD1W; CMH15; HEL114; MV; MVCL; VCL