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  1. Link

    vimentin-antibody-ln-6-ab230171.pdf

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Validated using a knockout cell line

Anti-Vimentin antibody [LN-6] (ab230171)

  • Datasheet
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Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [LN-6] (ab230171)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)

Key features and details

  • Mouse monoclonal [LN-6] to Vimentin
  • Suitable for: IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgM

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Overview

  • Product name

    Anti-Vimentin antibody [LN-6]
    See all Vimentin primary antibodies
  • Description

    Mouse monoclonal [LN-6] to Vimentin
  • Host species

    Mouse
  • Tested applications

    Suitable for: IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Human
  • Immunogen

    Tissue, cells or virus. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: HeLa cells, HAP1 and HeLa Wild-type cells. IHC-P: Human kidney (FFPE).
  • General notes

    Knockout and wild type cell lines were used to confirm the specificity of this antibody in ICC/IF.

    This antibody clone is manufactured by Abcam. If you require a custom buffer formulation or conjugation for your experiments, please contact orders@abcam.com.

    The Life Science industry has been in the grips of a reproducibility crisis for a number of years. Abcam is leading the way in addressing this with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation. 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, along with publications, customer reviews and Q&As

Properties

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    Preservative: 0.02% Sodium azide
    Constituents: PBS, 6.97% L-Arginine
  • Concentration information loading...
  • Clonality

    Monoclonal
  • Clone number

    LN-6
  • Isotype

    IgM
  • Research areas

    • Neuroscience
    • Cell Type Marker
    • Neural Stem Cell marker
    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Intermediate Filaments
    • Class III
    • Vimentin
    • Neuroscience
    • Neurology process
    • Neurogenesis
    • Stem Cells
    • Lineage Markers
    • Endoderm
    • Stem Cells
    • Neural Stem Cells
    • Intracellular
    • Cancer
    • Tumor biomarkers
    • Other
    • Neuroscience
    • Neurology process
    • Neuroregeneration
    • Neuroregeneration
    • Developmental Biology
    • Lineage specification
    • Ectoderm
    • Neuroscience
    • Development
    • Neuroscience
    • Processes

Associated products

  • Alternative Versions

    • Anti-alpha smooth muscle Actin antibody [EPR5368] (ab124964)
    • Anti-Vimentin antibody (ab24525)
    • Anti-alpha smooth muscle Actin (acetyl E3) + ACTG2 (acetyl E3) antibody [E184] (ab32575)
    • Anti-Vimentin antibody - Cytoskeleton Marker (ab45939)
    • Anti-alpha smooth muscle Actin antibody (ab5694)
    • Anti-Vimentin antibody [V9] - Cytoskeleton Marker (ab8069)
    • Anti-Vimentin antibody [RV202] - Cytoskeleton Marker (ab8978)
    • Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547)
  • Compatible Secondaries

    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) (ab150113)
    • Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab230171 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
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 µg/ml.
Notes
IHC-P
Use a concentration of 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF
Use a concentration of 1 µg/ml.

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.
  • Target information above from: UniProt accession P08670 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 7431 Human
    • Omim: 193060 Human
    • SwissProt: P08670 Human
    • Unigene: 455493 Human
    • Unigene: 691131 Human
    • 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

    • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)
      Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)

      ab230171 staining Vimentin (colored green) in wild-type HAP1 cells (top panel) and VIM knockout HAP1 cells (bottom panel). The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab230171 at 1μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution overnight at 4ºC. Cells were then incubated with ab150077, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labelled with DAPI (shown in blue).

      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).

    • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)
      Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)

      ab230171 staining Vimentin in HeLa-VIM cells. The cells were fixed with 4% paraformaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated overnight at 4°C with ab230171 at 1µg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control. Cells were then incubated with ab150121, Goat polyclonal Secondary Antibody to Mouse IgM - mu chain (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150080, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolour red). Nuclear DNA was labelled with DAPI (shown in blue).

      Also suitable in cells fixed with 100% methanol (5 min).

      Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [LN-6] (ab230171)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [LN-6] (ab230171)

      IHC image of Vimentin staining in a section of formalin-fixed paraffin-embedded normal human kidney* performed on a Leica BONDTM system using the standard protocol F.

      The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH 6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab230171, 1 µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with hematoxylin and mounted with DPX. The inset secondary-only 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

    • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)
      Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [LN-6] (ab230171)

      ab230171 staining Vimentin in HeLa (Human epithelial cell line from cervix adenocarcinoma) cells.

      The cells were fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 5 mins and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab230171 at 1 μg/ml and ab6046, Rabbit polyclonal to beta Tubulin - Loading Control, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) at 1/1000 dilution (shown in green) and ab150084, Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 594) at 1/1000 dilution (shown in pseudocolor red). Nuclear DNA was labeled with DAPI (shown in blue).

    Protocols

    To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.

    Click here to view the general protocols

    Datasheets and documents

    • Datasheet download

      Download

    References (1)

    Publishing research using ab230171? Please let us know so that we can cite the reference in this datasheet.

    ab230171 has been referenced in 1 publication.

    • Stathopoulos E  et al. LN-6: a monoclonal antibody to vimentin expressed in non-hematopoietic mesenchymal cells and derived tumors and reactive in B5-fixed, paraffin-embedded tissues. J Histochem Cytochem 37:1363-70 (1989). IP, IHC-P, IHC-Fr, ICC/IF ; Human . PubMed: 2671152

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