Human/Rat Vinculin Antibody

  
  • Species Reactivity
    Human, Rat
  • Specificity
    Detects recombinant human Vinculin in direct ELISAs and human and rat Vinculin in Western blots.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    E. coli-derived recombinant human Vinculin
    Lys1020-Gln1134
    Accession # P18206
  • Formulation
    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.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human and Rat Vinculin by Western Blot. Western blot shows lysates of HeLa human cervical epithelial carcinoma cell line and rat heart tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human/Rat Vinculin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6896) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). A specific band was detected for Vinculin at approximately 120 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
Vinculin in Human Uterus. Vinculin was detected in immersion fixed paraffin-embedded sections of human uterus using Sheep Anti-Human/Rat Vinculin Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6896) at 3 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-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counter­stained with hematoxylin (blue). vView our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Reconstitute at 0.2 mg/mL in sterile PBS.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & 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.
Background: Vinculin

VCL (Vinculin; from the Latin vinculum denoting a union or connection) is a 115-120 kDa cytoplasmic member of the vinculin/ alpha -catenin family of proteins. The VCL gene is ubiquitously expressed and generates a series of globular domains whose N-and C-terminal globular regions self-associate to generate an inactive molecule. When at least two ligands bind to the molecule, it opens up, allowing for the formation of multiple complexes that impact both signaling and cell structure. Among other things, VCL is known to regulate the expression of E-cadherin, and to connect actin to integrins via talin. Human VCL is 1134 amino acids (aa) in length, and at this size, is commonly known as metavinculin. It contains an 835 aa N-terminal globular head that binds talin, beta - and alpha -catenin, followed by a Pro-rich linker segment (aa 836-878) and a 27-29 kDa C-terminal tail that binds lipids plus paxillin and PKC alpha (aa 879-1134). At least seven utilized phosphorylation sites exist. There are three potential splice variants. One shows a deletion of aa 916-983 and is called vinculin. A second contains an alternative start site at Met74, and again possesses a deletion of aa 916-983. A third shows a 34 aa substitution for aa 262‑1134. Over the range of aa used as the immunogen for this reagent (aa 1020-1134 in metavinculin; 952-1066 in vinculin), human and mouse VCL are identical in aa sequence.

  • Long Name:
    Metavinculin
  • Entrez Gene IDs:
    7414 (Human); 22330 (Mouse); 305679 (Rat)
  • Alternate Names:
    CMD1W; CMH15; metavinculin; MVCL; VCL; Vinculin
Related Research Areas
Cell and Tissue Staining Kits
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Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

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Immunohistochemistry Reagents
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Isotype Controls
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Normal Sheep IgG Control

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Secondary Antibodies
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Sheep IgG HRP-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

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Sheep IgG (H+L) PE-conjugated Antibody

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Donkey Anti-Sheep IgG Biotinylated Antibody

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Sheep IgG (H+L) APC-conjugated Antibody

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Sheep IgG VisUCyte HRP Polymer Antibody

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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

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