Vinculin Antibody Cat. No.: 7809 Vinculin Antibody Figure 2 Western Blot Validation in Human, Mouse and Rat Cell Lines Figure 3 Western Blot Validation in PC3 Cell Lysate Loading: 15 μg of lysates per lane. Antibodies: Vinculin Loading: 15 μg of lysates per lane. Antibodies: Vinculin 7809 (1 μg/mL), 1h incubation at RT in 5% NFDM/TBST. 7809 (Lane 1: 0.5 μg/mL and Lane 2: 1 μg/mL), 1h Secondary: Goat anti-rabbit IgG HRP conjugate at 1:10000 incubation at RT in 5% NFDM/TBST. Secondary: Goat anti- dilution. rabbit IgG HRP conjugate at 1:10000 dilution. Figure 4 Immunocytochemistry Validation of Vinculin in Jurkat Cells Figure 5 Immunofluorescence Validation of Vinculin in Immunocytochemical analysis of Jurkat cells using anti- Jurkat Cells Vinculin antibody (7809) at 5 μg/ml. Cells was fixed with Immunofluorescent analysis of 4% paraformaldehyde- formaldehyde and blocked with 10% serum for 1 h at RT; fixed Jurkat cells labeling Vinculin with 7809 at 20 μg/mL, antigen retrieval was by heat mediation with a citrate followed by goat anti-rabbit IgG secondary antibody at buffer (pH6). Samples were incubated with primary 1/500 dilution (green) and DAPI staining (blue). antibody overnight at 4˚C. A goat anti-rabbit IgG H&L (HRP) at 1/250 was used as secondary. Counter stained with Hematoxylin. Specifications September 23, 2021 1 https://www.prosci-inc.com/vinculin-antibody-7809.html HOST SPECIES: Rabbit SPECIES REACTIVITY: Human, Mouse, Rat HOMOLOGY: Predicted species reactivity based on immunogen sequence: Pig: (100%) Anti-Vinculin antibody (7809) was raised against a peptide corresponding to 23 amino IMMUNOGEN: acids near the center of human Vinculin. < TESTED APPLICATIONS: ELISA, ICC, IF, WB WB: 0.5-2 μg/mL; ICC: 5 μg/mL; IF: 20 μg/mL. APPLICATIONS: Antibody validated: Western Blot in human, mouse and rat samples; Immunocytochemistry in human samples; Immunofluorescence in human samples. All other applications and species not yet tested. Vinculin antibody is human, mouse and rat reactive. At least three isoforms of Vinculin are SPECIFICITY: known to exist. POSITIVE CONTROL: 1) Cat. No. 1202 - A431 Cell Lysate 2) Cat. No. 1201 - HeLa Cell Lysate 3) Cat. No. 1282 - 3T3/NIH Cell Lysate Predicted: 124kD PREDICTED MOLECULAR WEIGHT: Observed: 124kD Advanced Validation Independent Antibody Validation in Cell lines (Figure 2) shows similar Vinculin expression profile in human, mouse and rat cell lines detected by three independent anti- Vinculin antibodies that recognize different epitopes, 7489 against C-terminus domain, VALIDATION: 7807 against N-terminus domain and 7809 against central domain. Vinculin proteins are detected in the most tested cell lines at different expression levels by the three independent antibodies. Human Vinculin has 3 isoforms, including isoform 2 (1134aa, 124kD), isoform 1 (1066aa, 117kD) and isoform 3 (222aa, 25kD). Mouse Vinculin has one isoform (1066aa, 117kD) and ISOFORMS: Rat Vinculin also has one isoform (1066aa, 117kD). 7809 can detect human, mouse and rat. Properties PURIFICATION: Vinculin antibody is affinity chromatography purified via peptide column. CLONALITY: Polyclonal ISOTYPE: IgG CONJUGATE: Unconjugated PHYSICAL STATE: Liquid BUFFER: Vinculin antibody is supplied in PBS containing 0.02% sodium azide. September 23, 2021 2 https://www.prosci-inc.com/vinculin-antibody-7809.html CONCENTRATION: 1 mg/mL Vinculin antibody can be stored at 4˚C for three months and -20˚C, stable for up to one STORAGE CONDITIONS: year. Additional Info OFFICIAL SYMBOL: VCL ALTERNATE NAMES: Vinculin Antibody: MV, MVCL, CMD1W, CMH15, HEL114, Vinculin, Metavinculin, MV ACCESSION NO.: NP_003364 PROTEIN GI NO.: 4507877 GENE ID: 7414 USER NOTE: Optimal dilutions for each application to be determined by the researcher. Background and References Vinculin is a cytoskeletal protein that plays an important role in the regulation of focal adhesions and embryonic development (1). Three structural vinculin domains include an amino-terminal head, a short flexible proline-rich region and a carboxy-terminal tail (2). BACKGROUND: Expression of vinculin were shown to be affected by the level of actin expression (2,3). Vinculin deficiencies are associated with a decrease in cell adhesion and an increase in cell motility, suggesting a possible role in metastatic growth (4). Defects in VCL are the cause of cardiomyopathy dilated type 1W (CMD1W) (5). REFERENCES: 1) Burridge et al. Annu. Rev. Cell Biol.1988; 4:487-525. 2) Gilmore et al. J. Cell Sci. 1992; 103:719-31. 3) Deakin et al. PLoS One 2012; 7:e37990. 4) Goldmann et al. Cell Biol. Int. 2013; Feb 1. CITATIONS 1)Wang et al. AMPK is involved in the differential neonatal performance of chicks hatching CITATIONS: at different time. Gen Comp Endocrinol. 2016;228:53-59. PMID: 26873631 ANTIBODIES FOR RESEARCH USE ONLY. For additional information, visit ProSci's Terms & Conditions Page. September 23, 2021 3 https://www.prosci-inc.com/vinculin-antibody-7809.html.