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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

  • Datasheet
  • SDS
  • Certificate of Compliance
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Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
  • Flow Cytometry - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
  • Flow Cytometry - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
  • Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Alexa Fluor® 647 Rabbit monoclonal [EPR3776] to Vimentin - Cytoskeleton Marker
  • Suitable for: Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human
  • Conjugation: Alexa Fluor® 647. Ex: 652nm, Em: 668nm

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Overview

  • Product name

    Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker
    See all Vimentin primary antibodies
  • Description

    Alexa Fluor® 647 Rabbit monoclonal [EPR3776] to Vimentin - Cytoskeleton Marker
  • Host species

    Rabbit
  • Conjugation

    Alexa Fluor® 647. Ex: 652nm, Em: 668nm
  • Tested Applications & Species

    Application Species
    Flow Cyt
    Mouse
    Human
    ICC/IF
    Mouse
    Human
    See all applications and species data
  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

    • ICC/IF: NIH 3T3 and wildtype HAP1 cells. Flow Cyt: NIH 3T3 cells and wildtype HAP1 cells.
  • General notes

     

     

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

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. Avoid freeze / thaw cycle. Store In the Dark.
  • Dissociation constant (KD)

    KD = 1.10 x 10 -10 M
    Learn more about KD
  • Storage buffer

    pH: 7.40
    Preservative: 0.02% Sodium azide
    Constituents: PBS, 30% Glycerol (glycerin, glycerine), 1% BSA
  • Concentration information loading...
  • Purity

    Immunogen affinity purified
  • Clonality

    Monoclonal
  • Clone number

    EPR3776
  • Isotype

    IgG
  • 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

Associated products

  • Alternative Versions

    • Alexa Fluor® 594 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab154207)
    • Alexa Fluor® 488 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab185030)
    • Anti-Vimentin antibody [EPR3776] - BSA and Azide free (ab193555)
    • HRP Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194718)
    • Alexa Fluor® 568 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab202504)
    • Alexa Fluor® 555 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab203428)
    • PE Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab209446)
    • Alexa Fluor® 405 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab210152)
    • Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab92547)
  • Isotype control

    • Alexa Fluor® 647 Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab199093)
  • Recombinant Protein

    • Recombinant Human Vimentin protein (ab73843)
  • Related Products

    • Human Vimentin Profiling ELISA Kit (ab173190)

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab194719 in the following tested applications.

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

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
Flow Cyt
Mouse
Human
ICC/IF
Mouse
Human
All applications
Rat
Application Abreviews Notes
Flow Cyt
1/500.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody.

 

ICC/IF
1/100 - 1/1000.
Notes
Flow Cyt
1/500.

ab199093 - Rabbit monoclonal IgG (Alexa Fluor® 647), is suitable for use as an isotype control with this antibody.

 

ICC/IF
1/100 - 1/1000.

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
    • Entrez Gene: 22352 Mouse
    • Entrez Gene: 81818 Rat
    • Omim: 193060 Human
    • SwissProt: P08670 Human
    • SwissProt: P20152 Mouse
    • SwissProt: P31000 Rat
    • Unigene: 455493 Human
    • Unigene: 691131 Human
    • Unigene: 268000 Mouse
    • Unigene: 2710 Rat
    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

  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

    ab194719 staining Vimentin in wild-type HAP1 cells (top panel) and Vimentin 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 ab194719 at 1/1000 dilution (shown in red) and ab195887 at 1/250 dilution (shown in green) overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

  • Flow Cytometry - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
    Flow Cytometry - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

    Overlay histogram showing HAP1 wildtype (green line) and HAP1-VIM  knockout cells (red line) stained with ab194719. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab194719, 0.01µg/ml dilution) for 30 min at 22°C.

    A rabbit monoclonal IgG isotype control antibody  (ab199093) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-VIM knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity).

    Acquisition of >5,000 events were collected using a 40 mW Yellow/Green laser (640nm) and 670/14 bandpass filter.

    This antibody can also be used in HAP1 cells fixed with 4% formaldehyde (10 min), permeabilized with 0.1% PBS-Triton X-100 for 15 min under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
    Immunocytochemistry/ Immunofluorescence - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

    ab194719 staining Vimentin in NIH3T3 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab194719 at 1/100 dilution(shown in red) and ab195887, Mouse monoclonal [DM1A] to alpha Tubulin (Alexa Fluor® 488, shown in green) at 2µg/ml overnight at +4°C. Nuclear DNA was labelled in blue with DAPI.

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

  • Flow Cytometry - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
    Flow Cytometry - Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

    Overlay histogram showing NIH3T3 cells stained with ab194719 (red line). The cells were fixed with 4% formaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab194719, 1/500 dilution) for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) Alexa Fluor® 647 used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a solid-state 25mW red diode laser (635 nm) and 675/30 bandpass filter.

    This antibody gave a positive signal in NIH3T3 fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

  • Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)
    Alexa Fluor® 647 Anti-Vimentin antibody [EPR3776] - Cytoskeleton Marker (ab194719)

Protocols

  • Flow cytometry protocols
  • Immunocytochemistry & immunofluorescence protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

Certificate of Compliance

To download a Certificate of Compliance, please enter your Lot number below:

References (2)

Publishing research using ab194719? Please let us know so that we can cite the reference in this datasheet.

ab194719 has been referenced in 2 publications.

  • Wei F  et al. Plasma endothelial cells-derived extracellular vesicles promote wound healing in diabetes through YAP and the PI3K/Akt/mTOR pathway. Aging (Albany NY) 12:12002-12018 (2020). PubMed: 32570219
  • Sakkou M  et al. Mesenchymal TNFR2 promotes the development of polyarthritis and comorbid heart valve stenosis. JCI Insight 3:N/A (2018). PubMed: 29618659

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