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Validated using a knockout cell lineRabMAb

Anti-Vimentin antibody [SP20], prediluted (ab27608)

Submit a review Q&A (1)References (10)
Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)
  • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20], prediluted (ab27608)
  • Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20], prediluted (ab27608)
  • Flow Cytometry - Anti-Vimentin antibody [SP20], prediluted (ab27608)

Key features and details

  • Rabbit monoclonal [SP20] to Vimentin, prediluted
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Human
  • Isotype: IgG

Overview

  • Product name

    Anti-Vimentin antibody [SP20], prediluted
    See all Vimentin primary antibodies

  • Description

    Rabbit monoclonal [SP20] to Vimentin, prediluted

  • Host species

    Rabbit

  • Tested applications

    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein corresponding to Human Vimentin.

  • Positive control

    • WB: HeLa, U20S and human tonsil cell lysates. IHC-P: Human melanoma. Flow Cyt: HeLa cells.

  • General notes

    This product is FOR RESEARCH USE ONLY. For commercial use, please contact partnerships@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

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab27608 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
Flow Cyt
1/1.

Use neat.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
IHC-P
Use at an assay dependent concentration. Do not perform antigen retrieval.
ICC/IF
Use at an assay dependent concentration.
Notes
Flow Cyt
1/1.

Use neat.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
WB
Use at an assay dependent concentration. Predicted molecular weight: 53 kDa.
IHC-P
Use at an assay dependent concentration. Do not perform antigen retrieval.
ICC/IF
Use at an assay dependent concentration.

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

    • 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 - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      All lanes : Anti-Vimentin antibody [SP20] (ab16700) at 1/100 dilution

      Lane 1 : U20S cell lysate
      Lane 2 : Human tonsil cell lysate
      Lane 3 : Wild-type HeLa cell lysate
      Lane 4 : VIM knockout HeLa cell lysate

      Lysates/proteins at 20 µg per lane.

      Performed under reducing conditions.

      Predicted band size: 53 kDa
      Observed band size: 53 kDa



      This data was developed using the same antibody clone in a different buffer formulation (ab16700).

      Lanes 1 - 4: Merged signal (red and green). Green - ab16700 observed at 53 kDa. Red - loading control, ab8245 observed at 37 kDa.

       ab16700 was shown to react with Vimentin in wild-type HeLa cells. Loss of signal was observed when knockout cell line ab255446 (knockout cell lysate ab263775) was used. Wild-type and Vimentin knockout samples were subjected to SDS-PAGE. ab16700 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 100 dilution and 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

       

       

    • Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Immunocytochemistry/ Immunofluorescence - Anti-Vimentin antibody [SP20], prediluted (ab27608)

      ab16700 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 ab16700 at 1/1000 dilution and ab195889 at 1/250 dilution (shown in pseudo-color red) overnight at +4°C. The cells were then incubated with ab150081 (Goat polyclonal Secondary Antibody to Rabbit IgG - H&L (Alexa Fluor® 488)) at 1/1000 dilution for 1 hour. Nuclear DNA was labelled in blue with DAPI.

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

      This data was developed using the undiluted version of this antibody (ab16700).

    • Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Western blot - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Anti-MDM2 antibody [4B11] (ab16900) at 1/100 dilution + HeLa cell lysate

      Predicted band size: 53 kDa
      Observed band size: 53 kDa



      This image refers to the non-prediluted version of this product (ab16900)
    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Human melanoma stained with prediluted ab27608.
    • Flow Cytometry - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Flow Cytometry - Anti-Vimentin antibody [SP20], prediluted (ab27608)
      Overlay histogram showing HeLa cells stained with ab27608 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween 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 (ab27608, neat) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Unlabelled sample (blue line). Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

    References (10)

    Publishing research using ab27608? Please let us know so that we can cite the reference in this datasheet.

    ab27608 has been referenced in 10 publications.

    • Medzikovic L  et al. Nuclear Receptor Nur77 Controls Cardiac Fibrosis through Distinct Actions on Fibroblasts and Cardiomyocytes. Int J Mol Sci 22:N/A (2021). PubMed: 33562500
    • Liu Y  et al. FOXA2-Interacting FOXP2 Prevents Epithelial-Mesenchymal Transition of Breast Cancer Cells by Stimulating E-Cadherin and PHF2 Transcription. Front Oncol 11:605025 (2021). PubMed: 33718155
    • Nizyaeva NV  et al. Ultrastructural and Immunohistochemical Features of Telocytes in Placental Villi in Preeclampsia. Sci Rep 8:3453 (2018). IHC-P ; Human . PubMed: 29472628
    • Ye D  et al. WTX inhibits gastric cancer migration through the reversal of epithelial-mesenchymal transition. Oncol Lett 16:4970-4976 (2018). PubMed: 30250562
    • Zangari J  et al. Rapid decay of engulfed extracellular miRNA by XRN1 exonuclease promotes transient epithelial-mesenchymal transition. Nucleic Acids Res 45:4131-4141 (2017). WB ; Human . PubMed: 27994032
    • Jennings AR & Carroll WM Oligodendrocyte Lineage Cells in Chronic Demyelination of Multiple Sclerosis Optic Nerve. Brain Pathol 25:517-30 (2015). PubMed: 25175564
    • Walker E  et al. Microscopic detection of quenched activity-based optical imaging probes using an antibody detection system: localizing protease activity. Mol Imaging Biol 16:608-18 (2014). PubMed: 24705781
    • Yang C  et al. FOXM1 promotes the epithelial to mesenchymal transition by stimulating the transcription of Slug in human breast cancer. Cancer Lett 340:104-12 (2013). WB ; Human . PubMed: 23856032
    • Ahmad A  et al. Phosphoglucose isomerase/autocrine motility factor mediates epithelial-mesenchymal transition regulated by miR-200 in breast cancer cells. Cancer Res 71:3400-9 (2011). WB ; Human . PubMed: 21389093
    • Mirabet V  et al. Long-term storage in liquid nitrogen does not affect cell viability in cardiac valve allografts. Cryobiology 57:113-21 (2008). IHC-Fr . PubMed: 18703039

    Customer reviews and Q&As

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    Submit a review Submit a question

    Question

    What does 'pre-diluted' antibody mean? As an example AB27608 (Vimentin). It says "use at an assay dependent dilution?" So, should I furthere dilute the antibody to get myf inal concentration or is the antibody already diluted for use. Also, what is the total protein concentration in AB27608?

    Read More
    Answer

    Thank you for contacting us. The antibody is ready to use and should not be further diluted.Because the antibody is from tissue culture supernatant and we do not routinely determine the concentration.

    Read More

    Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
    For licensing inquiries, please contact partnerships@abcam.com