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Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)
![Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)](/ps/datasheet/images/ab18058_1.jpg)
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Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)
![Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)](/ps/datasheet/images/ab18058_2.jpg)
(enlarge)
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Western blot - Vinculin antibody [SPM227] (ab18058)
![Western blot - Vinculin antibody [SPM227] (ab18058)](/ps/datasheet/images/18/ab18058/Vinculin-Primary-antibodies-ab18058-3.jpg)
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Product Name
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Vinculin antibody [SPM227]
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See all Vinculin antibodies (12)...
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Product type
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Primary antibodies
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Description
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Mouse monoclonal [SPM227] to Vinculin
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Immunogen
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Full length native protein (semi-purified) (Human).
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Reacts with
(species key)
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Hu, Ms, Hm
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Tested applications
(see key)
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ICC/IF, IHC-P, IP, WB
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Abreviews
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Application notes
(see key)
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Recommended dilutions ICC/IF: 1/200 (see Abreview). IHC-P: 1/25 for 30 min at RT. Staining of formalin-fixed tissues requires boiling tissue sections in 10mM citrate buffer, pH 6.0 for 10 min followed by cooling at RT for 20 min. IP: Use at 2µg/mg of lysate. WB: Use at a concentration of 1 - 2 ug/ml for 2hrs at RT. Predicted molecular weight: 130kDa (vinculin) and 150kDa (meta-vinculin)
Not tested in other applications. Optimal dilutions/concentrations should be determined by the end user.
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Positive control
(see definition)
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HeLa cells, skin.
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Cellular localization
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Cytoplasm; cytoskeleton. Cell junction; adherens junction; cytoplasmic side. Cytoplasmic face of adhesion plaques.
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Research areas
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Cardiovascular >> Heart >> Contractility >> Contractile Proteins >> Actins Cardiovascular >> Heart >> Cardiac arrhythmias Signal Transduction >> Cytoskeleton / ECM >> Cytoskeleton >> Microfilaments >> Actin etc >> Actin Binding Proteins
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Relevance
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Vinculin is a cytoskeletal protein associated with the cytoplasmic faces of both cell-cell and cell extracellular matrix adherens type junctions. It functions as one of several interacting proteins involved in anchoring F-actin to the membrane. The sequences of chicken, nematode and human vinculin have been determined. It has been shown that a sequence of molecular interactions might be involved in the transmembrane assembly of adhesion plaques. In the assembly of adhesion plaques, the beta subunit of integrin binds to talin. Talin binds to vinculin that interacts with alpha actinin and possibly with itself. Since alpha actinin binds to and cross links actin filaments, vinculin represents a key element in the transmembrane linkage of the extracellular matrix to the cytoplasmic microfilament system.
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Database links
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The links below go to external sites and will open in a new browser window
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Raised in
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Mouse
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Clonality
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Monoclonal
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Clone number
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SPM227
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Isotype
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IgG1
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Light chain type
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kappa
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Purity
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Protein G purified
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Storage buffer
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Preservative: Sodium Azide Constituents: BSA, 10mM PBS, pH 7.4 Material safety datasheet (MSDS) for this product: Sodium Azide MSDS
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Purification notes
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Purified from ascites fluid by protein G
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Form
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Liquid
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Concentration
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0.200 mg/ml
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Storage instructions
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Store at +4°C.
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At Abcam, we have one centralized database to hold all of our product information, so that everything we know about this Vinculin antibody [SPM227] is on this datasheet. But please do contact us if you would like any reassurance! |
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See below for Vinculin antibody [SPM227] images, references, products related to ab18058 and other tools.
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Vinculin antibody [SPM227] images:
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Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)
ab18058 at 1/200 staining Chinese Hamster Ovary cells by ICC/IF. The cells were permeabilized with 100% methanol and then blocked with 1% BSA/ 4% goat serum prior to incubation with the antibody for 30 minutes. A rhodamine conjugated goat antibody was used as the secondary. This image is courtesy of an anonymous Abreview
See Abreview
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Immunocytochemistry/ Immunofluorescence - Vinculin antibody [SPM227] (ab18058)
ab18058 staining mouse 3T3 cells by ICC/IF. Cells were PFA fixed and permeabilized in Triton X-100 prior to blocking in 1% BSA for 30 minutes at 25°C. The primary antibody was diluted 1/100 and incubated with the sample for 1 hour at 25°C, ab6785, diluted 1/100, was used as the secondary antibody. This image is courtesy of an Abreview submitted by Mr John Maloney
See Abreview
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Western blot - Vinculin antibody [SPM227] (ab18058)
Vinculin antibody [SPM227] (ab18058) at 1/1000 dilution + Whole cell lysate prepared from human lung cells at 40 µg
Secondary HRP conjugated goat polyclonal to mouse IgG at 1/1000 dilution
Predicted band size : 130 kDa Observed band size : 117 kDa (why is the actual band size different from the predicted?)
The image is a courtsey of an anonymous abreview.
See Abreview
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)-Vinculin antibody [SPM227](ab18058)
Ab18058 staining human normal prostate. Staining is localised to the nucleus and cytoplasm. Left panel: with primary antibody at 1 ug/ml. Right panel: isotype control. Sections were stained using an automated system DAKO Autostainer Plus , at room temperature. Sections were rehydrated and antigen retrieved with the Dako 3-in-1 AR buffer citrate pH 6.0 in a DAKO PT Link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 minutes. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS), then incubated with primary antibody for 20 minutes, and detected with Dako Envision Flex amplification kit for 30 minutes. Colorimetric detection was completed with diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that for manual staining we recommend to optimize the primary antibody concentration and incubation time (overnight incubation), and amplification may be required.
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Western blot - Vinculin antibody [SPM227] (ab18058)
Predicted band size : 130 kDa
Lanes: 1, 4, 7 - Hamster Lung; 2, 5, 8 - K562; 3, 6, 9 - CHO Lanes: 1-3: 4oC (1 freeze/thaw); 4-6: 4oC (2 freeze/thaws); 7-9 : 4oC (4 freeze/thaws)
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References for Vinculin antibody [SPM227] (ab18058)
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This product has been used in: (two most recent references)
Raya A et al. Disease-corrected haematopoietic progenitors from Fanconi anaemia induced pluripotent stem cells. Nature 460:53-9 (2009). WB; Human. PubMed: 19483674
Hahn CK et al. Proteomic and genetic approaches identify Syk as an AML target. Cancer Cell 16:281-94 (2009). WB; Human. PubMed: 19800574
If you publish research using ab18058 please let us know so that we can cite the reference on this datasheet.
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Search PubMed (MEDLINE) for references to Vinculin
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Vinculin antibody [SPM227] - more information
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Customer reviews (feedback) regarding Vinculin antibody [SPM227] |
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Vinculin antibody [SPM227] - related products:
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Vinculin antibody [SPM227] - other tools:
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Contact Abcam with a Technical Enquiry about Vinculin antibody [SPM227] |
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All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"