Validated using a knockout cell line

HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)

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Western blot - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
  • Western blot - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)

Key features and details

  • HRP Mouse monoclonal [V9] to Vimentin - Cytoskeleton Marker
  • Suitable for: IHC-P, WB
  • Knockout validated
  • Reacts with: Human
  • Conjugation: HRP
  • Isotype: IgG1

Overview

  • Product name

    HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker
    See all Vimentin primary antibodies

  • Description

    HRP Mouse monoclonal [V9] to Vimentin - Cytoskeleton Marker

  • Host species

    Mouse

  • Conjugation

    HRP

  • Tested applications

    Suitable for: IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human
    Predicted to work with: Rat, Cow

  • Positive control

    • WB: HeLa and MOLT4 whole cell lysates. IHC-P: normal human colon tissue.

  • General notes

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

Properties

Applications

Our Abpromise guarantee covers the use of ab196602 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Application Abreviews Notes
IHC-P 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
WB 1/5000. Detects a band of approximately 57 kDa (predicted molecular weight: 54 kDa).

Target

  • Function

    Vimentins are class-III intermediate filaments found in various non-epithelial cells, especially mesenchymal cells. Vimentin is attached to the nucleus, endoplasmic reticulum, and mitochondria, either laterally or terminally.
    Involved with LARP6 in the stabilization of type I collagen mRNAs for CO1A1 and CO1A2.

  • Tissue specificity

    Highly expressed in fibroblasts, some expression in T- and B-lymphocytes, and little or no expression in Burkitt's lymphoma cell lines. Expressed in many hormone-independent mammary carcinoma cell lines.

  • Involvement in disease

    Cataract 30

  • Sequence similarities

    Belongs to the intermediate filament family.

  • Domain

    The central alpha-helical coiled-coil rod region mediates elementary homodimerization.
    The [IL]-x-C-x-x-[DE] motif is a proposed target motif for cysteine S-nitrosylation mediated by the iNOS-S100A8/A9 transnitrosylase complex.

  • Post-translational
    modifications

    Filament disassembly during mitosis is promoted by phosphorylation at Ser-55 as well as by nestin (By similarity). One of the most prominent phosphoproteins in various cells of mesenchymal origin. Phosphorylation is enhanced during cell division, at which time vimentin filaments are significantly reorganized. Phosphorylation by PKN1 inhibits the formation of filaments. Phosphorylated at Ser-56 by CDK5 during neutrophil secretion in the cytoplasm. Phosphorylated by STK33.
    O-glycosylated during cytokinesis at sites identical or close to phosphorylation sites, this interferes with the phosphorylation status.
    S-nitrosylation is induced by interferon-gamma and oxidatively-modified low-densitity lipoprotein (LDL(ox)) possibly implicating the iNOS-S100A8/9 transnitrosylase complex.

  • Cellular localization

    Cytoplasm.
  • Information by UniProt

  • Database links

    see all

  • Form

    Vimentin is found in connective tissue and in the cytoskeleton.

  • Alternative names

    • CTRCT30 antibody
    • Epididymis luminal protein 113 antibody
    • FLJ36605 antibody
    • HEL113 antibody
    • VIM antibody
    • VIME_HUMAN antibody
    • Vimentin antibody
    see all

Images

  • Western blot - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
    Western blot - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
    All lanes : HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602) at 1/5000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : VIM (Vimentin) knockout HAP1 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 54 kDa


    Exposure time: 1 minute


    ab196602 was shown to specifically react with Vimentin in wild-type HAP1 cells as signal was lost in VIM (Vimentin) knockout cells. Wild-type and VIM (Vimentin) knockout samples were subjected to SDS-PAGE. Ab196602 and ab184095 (Mouse monoclonal [mAbcam 9484] to GAPDH - Loading Control (Alexa Fluor® 680) loading control) were incubated overnight at 4°C at 1/5000 dilution and 1/1000 dilution respectively. The loading control was imaged using the Licor Odyssey CLx prior to blots being developed with ECL technique.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)

    IHC image of Vimentin staining in a section of formalin-fixed paraffin-embedded normal human colon tissue*, performed on a Leica BOND. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20mins. The section was then incubated with ab196602 at 1/100 dilution, for 15 mins at room temperature. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay without primary antibody.

    For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

    *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

  • Western blot - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
    Western blot - HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602)
    All lanes : HRP Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab196602) at 1/5000 dilution

    Lane 1 : HeLa whole cell lysate (ab150035)
    Lane 2 : MOLT4 (Human acute lymphoblastic leukemia cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 54 kDa
    Observed band size: 57 kDa
    why is the actual band size different from the predicted?


    Exposure time: 8 seconds


    This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 2% Bovine Serum Albumin before being incubated with ab196602 overnight at 4°C. Antibody binding was visualised using ECL development solution ab133406.

References (0)

Publishing research using ab196602? Please let us know so that we can cite the reference in this datasheet.

ab196602 has not yet been referenced specifically in any publications.

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