Vinculin Antibody (728526) [Unconjugated]

Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and HepG2 human hepatocellular carcinoma cell line. PVDF membrane was probed with 2 µg/mL of Mouse Anti-Human Vinculin Monoclonal Antibody (Catalog # MAB6896) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF007). A specific band was detected for Vinculin at approximately 124 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.

Simple Western lane view shows lysates of HeLa human cervical epithelial carcinoma cell line and PC‑3 human prostate cancer cell line, loaded at 0.2 mg/mL. A specific band was detected for Vinculin at approximately 122 kDa (as indicated) using 20 µg/mL of Mouse Anti-Human/Mouse/Rat Vinculin Monoclonal Antibody (Catalog # MAB6896). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.

Vinculin was detected in immersion fixed paraffin-embedded sections of human uterus using Mouse Anti-Human Vinculin Monoclonal Antibody (Catalog # MAB6896) at 15 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Mouse HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS002) and counterstained with hematoxylin (blue). Specific staining was localized to smooth muscle cells. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.

• Reactivity: Hu, Mu, Rt
• Applications: WB, Simple Western, IHC
• Clone: 728526
• Clonality: Monoclonal
• Host: Mouse
• Conjugate: Unconjugated
• Concentration: LYOPH

Vinculin Antibody (728526) [Unconjugated] Summary

• Immunogen: E. coli-derived recombinant human Vinculin
  Lys1020-Gln1134
  Accession # P18206
• Specificity: Detects human Vinculin in direct ELISAs and Western blots.
• Source: N/A
• Isotype: IgG2a
• Clonality: Monoclonal
• Host: Mouse
• Gene: VCL
• Purity Statement: Protein A or G purified from hybridoma culture supernatant

Applications/Dilutions

• Dilutions:
  • Western Blot 2 ug/mL
  • Simple Western 20 ug/mL
  • Immunohistochemistry 8-25 ug/mL
• Application Notes: In Simple Western only 10-15 uL of the recommended dilution is used per data point.

Packaging, Storage & Formulations

• Storage: Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
  • 12 months from date of receipt, -20 to -70 °C as supplied.
  • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
  • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
• Buffer: Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
• Preservative: No Preservative
• Concentration: LYOPH
• Reconstitution Instructions: Sterile PBS to a final concentration of 0.5 mg/mL.

Notes

This product is produced by and ships from R&D Systems, Inc., a Bio-Techne brand.

Alternate Names for Vinculin Antibody (728526) [Unconjugated]

• CMD1W
• CMH15
• metavinculin
• MVCL
• VCL
• Vinculin

Background

VCL (vinculin) is a 117 kDa actin binding cytoskeletal phosphoprotein found in adherens-type cell-cell and cell-matrix junctions. The 1066 amino acid (aa) human vinculin contains an N-terminal globular head (aa 2-835) with a talin interaction domain (aa 168-208) and three tandem repeats (aa 259-589), a pro-rich linker region (aa 826-878), and a C-terminal tail that facilitates phospholipid membrane insertion (aa 1003-1134). An 1134 aa form termed metavinculin contains 68 aa inserted after aa 915 and mainly occurs in cardiac, skeletal and smooth muscle. Human, mouse and rat vinculin share 100% aa sequence identity within the region used as an immunogen.

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Primary Antibodies are guaranteed for 1 year from date of receipt.

Publications for Vinculin Antibody (MAB6896)(5)

Publications using MAB6896

• CD Wiley, S Liu, C Limbad, AM Zawadzka, J Beck, M Demaria, R Artwood, F Alimirah, JA Lopez-Domi, C Kuehnemann, SR Danielson, N Basisty, HG Kasler, TR Oron, PY Desprez, SD Mooney, BW Gibson, B Schilling, J Campisi, P Kapahi SILAC Analysis Reveals Increased Secretion of Hemostasis-Related Factors by Senescent Cells Cell Rep, 2019;28(13):3329-3337.e5. 2019 [PMID: 31553904] (Western Blot, Human)
• A Schramm, P Mueller-Th, T Littmann, M Harloff, T Ozawa, J Schlossman Establishing a Split Luciferase Assay for Proteinkinase G (PKG) Interaction Studies Int J Mol Sci, 2018;19(4):. 2018 [PMID: 29649180] (Western Blot, Mouse)
• A Baradaran-, J Niesser, AD Balgi, K Choi, C Zimmerman, AP South, HJ Anderson, NC Strynadka, MB Bally, M Roberge Gentamicin B1 is a minor gentamicin component with major nonsense mutation suppression activity Proc. Natl. Acad. Sci. U.S.A, 2017;0(0):. 2017 [PMID: 28289221] (Western Blot Control, Mouse)
• Alireza Baradaran-, Aruna D Balgi, Carla Zimmerman, Kunho Choi, Fahimeh S Shidmooss, Jason S Tan, Célia Bergeaud, Alexandra Krause, Stéphane Flibotte, Yoko Shimizu, Hilary J Anderson, Vincent Mouly, Eric Jan, Tom Pfeifer, James B Jaquith, Michel Roberge Novel small molecules potentiate premature termination codon readthrough by aminoglycosides. Nucleic Acids Research, 2016;0(0):1362-4962. 2016 [PMID: 27407112] (Western Blot, Human)
• Naresh C Bal Increased reliance on muscle based thermogenesis upon acute minimization of brown adipose tissue function J Biol Chem, 2016;0(0):. 2016 [PMID: 27298322] (Western Blot, Mouse)

Reviews for Vinculin Antibody (MAB6896) (3)

Average Rating: 4.3

(Based on 3 reviews)

We have 3 reviews tested in 2 species: Human, Mouse.

reviewed by: Krishna Yalla Simple Western Human 08/27/2020

Summary

• Application: Simple Western
• Sample Tested: HCT116 cell line lysate
• Species: Human
• Lot: CFT00117091

Comments

• Comments: Ab titration on HCT116 cell lysate

reviewed by: Anonymous WB Human 10/16/2015

Summary

• Application: Western Blot
• Sample Tested: Hela whole cell lysate
• Species: Human
• Lot: CFTO0112081

Blocking

• Blocking Details: Blocker=BSA, Buffer= 1XTris Buffer Saline+0.1 % Tween 20, 2hrs at RT

Primary Anitbody

• Dilution Ratio: 1:500 over night at 4 degree Celsius

Secondary Antibody

• Secondary Description: anti-Mouse IgG, HRP
• Secondary Manufacturer Cat#: ab20043
• Secondary Concentration: 1:20,000

Details

• Detection Notes: 30 seconds exposure

Comments

• Comments: Yeah! it works

reviewed by: Asimina Kerimi Simple Western Mouse 09/23/2015

Summary

• Application: Simple Western
• Sample Tested: CaCo2 whole cell lysate
• Species: Mouse
• Lot: CFTO011508A

Blocking

• Blocking Details: 30 min incubation on Wes platform

Secondary Antibody

• Secondary Description: PS provided mouse HRP
• Secondary Manufacturer Cat#: n/a

Details

• Detection Notes: WES Protein Simple, Multi-image analysis used

Comments

• Comments: Cell lysate was collected in Bicine-Chaps Buffer

FAQs for Vinculin Antibody (MAB6896). (Showing 1 - 1 of 1 FAQs).

1. Do you have any 100 coda or bigger controls?
   • I can recommend a couple of high weight antibodies that are conserved in most samples. Please verify the presence of the protein before using as a loading control. PARP1 or PARP is approximately 113kDa, and you may view the products we offer to PARP using this link. Vinculin is approximately 116kDa, and you may view the products we offer to Vinculin using this link.

Bioinformatics

• Gene Symbol: VCL
• Uniprot:
  • P18206()