Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-Fascin antibody [EP5902] (ab126772)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP5902] to Fascin
  • Suitable for: WB, IP, IHC-P, Flow Cyt, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Fascin antibody [EP5902]
    See all Fascin primary antibodies

  • Description

    Rabbit monoclonal [EP5902] to Fascin

  • Host species

    Rabbit

  • Tested applications

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

  • Species reactivity

    Reacts with: Mouse, Rat, Human

  • Immunogen

    Recombinant fragment within Human Fascin aa 200-400. The exact sequence is proprietary.

  • Positive control

    • WB: HepG2, HAP1, HCT116, K562, SH SY5Y, U87-MG and K562 cell lysates. IHC-P: Human tonsil, glioma, kidney, colon and Hodgkin's lymphoma tissues. IP: HepG2 cell lysate. Flow Cyt: HeLa cells. ICC/IF: U87-MG cells.

  • General notes

     

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    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 -20°C. Stable for 12 months at -20°C.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 9% PBS, 40% Glycerol, 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...

  • Purity

    Tissue culture supernatant

  • Clonality

    Monoclonal

  • Clone number

    EP5902

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab126772 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
WB 1/10000 - 1/50000. Predicted molecular weight: 54 kDa.
IP 1/10 - 1/100.
IHC-P 1/250 - 1/500. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. (heat up to 98 degrees C, below boiling, and then let cool for 10-20 min.)
Flow Cyt 1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/500.

For unpurified use at 1/100 - 1/250

Target

  • Function

    Organizes filamentous actin into bundles with a minimum of 4.1:1 actin/fascin ratio. Plays a role in the organization of actin filament bundles and the formation of microspikes, membrane ruffles, and stress fibers. Important for the formation of a diverse set of cell protrusions, such as filopodia, and for cell motility and migration.

  • Tissue specificity

    Ubiquitous.

  • Sequence similarities

    Belongs to the fascin family.

  • Domain

    Composed of four beta-trefoil domains.

  • Post-translational
    modifications

    Phosphorylation on Ser-39 inhibits the actin-binding ability of fascin.

  • Cellular localization

    Cytoplasm > cytoskeleton. Cell projection > filopodium. Cell projection > invadopodium.
  • Information by UniProt

  • Database links

    see all

  • Alternative names

    • 55 kDa actin bundling protein antibody
    • 55 kDa actin-bundling protein antibody
    • Actin bundling protein antibody
    • actin bundling protein, 55-KD antibody
    • FAN 1 antibody
    • FAN1 antibody
    • Fascin 1 antibody
    • Fascin actin bundling protein 1 antibody
    • Fascin antibody
    • Fascin homolog 1 actin bundling protein (Strongylocentrotus purpuratus) antibody
    • Fascin homolog 1 antibody
    • Fascin, sea urchin, homolog of, 1 antibody
    • Fascin1 antibody
    • FLJ38511 antibody
    • FSCN 1 antibody
    • FSCN1 antibody
    • FSCN1_HUMAN antibody
    • HSN antibody
    • p55 antibody
    • Singed (Drosophila) like (sea urchin fascin homolog like) antibody
    • Singed drosophila homolog like antibody
    • Singed like (fascin homolog sea urchin) (Drosophila) antibody
    • Singed like (fascin homolog sea urchin) antibody
    • Singed like protein antibody
    • Singed, drosophila, homolog of antibody
    • Singed-like protein antibody
    • SNL antibody
    • Strongylocentrotus purpuratus antibody
    see all

Images

  • Western blot - Anti-Fascin antibody [EP5902] (ab126772)
    Western blot - Anti-Fascin antibody [EP5902] (ab126772)
    All lanes : Anti-Fascin antibody [EP5902] (ab126772) at 1/1000 dilution

    Lane 1 : Wild-type HCT116 cell lysate
    Lane 2 : FSCN1 knockout HCT116 cell lysate
    Lane 3 : SH-SY5Y cell lysate
    Lane 4 : K-562 cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) at 1/10000 dilution

    Predicted band size: 54 kDa
    Observed band size: 60 kDa
    why is the actual band size different from the predicted?



    Lanes 1-4: Merged signal (red and green). Green - ab126772 observed at 60 kDa. Red - loading control ab8245 observed at 36 kDa.

     ab126772 Anti-Fascin antibody [EP5902] was shown to specifically react with Fascin in wild-type HCT116 cells. Loss of signal was observed when knockout cell line ab266895 (knockout cell lysate ab257444) was used. Wild-type and Fascin knockout samples were subjected to SDS-PAGE. ab126772 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) were incubated overnight at 4°C at 1 in 1000 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-Fascin antibody [EP5902] (ab126772)
    Immunocytochemistry/ Immunofluorescence - Anti-Fascin antibody [EP5902] (ab126772)

    Immunocytochemistry/Immunofluorescence analysis of U87-MG (human glioblastoma) labelling Fascin with purified ab126772 at 1/500. Cells were fixed with 100% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (ab150077) at dilution of 1/1000 was used as the secondary antibody. Nuclei couterstained with DAPI (blue). 

    Secondary Only Control: PBS was used instead of the primary antibody as the negative control. 

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)

    ab126772, at 1/250 dilution, staining Fascin in paraffin-embedded Human Hodgkin's lymphoma tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Flow Cytometry - Anti-Fascin antibody [EP5902] (ab126772)
    Flow Cytometry - Anti-Fascin antibody [EP5902] (ab126772)
    Overlay histogram showing HeLa cells stained with ab126772 (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 (ab126772, 1/1000 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (0.1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
  • Immunoprecipitation - Anti-Fascin antibody [EP5902] (ab126772)
    Immunoprecipitation - Anti-Fascin antibody [EP5902] (ab126772)

    Purified ab126772 at 1/20 dilution (0.5µg) immunoprecipitating Fascin in HepG2 whole cell lysate.
    Lane 1 (input): HepG2 (Human hepatocellular carcinoma epithelial cell) whole cell lysate 10µg
    Lane 2 (+): ab126772 + HepG2 whole cell lysate.
    Lane 3 (-): Rabbit monoclonal IgG (ab172730) instead of ab126772 in HepG2 whole cell lysate.
    VeriBlot for IP Detection Reagent (HRP) (ab131366) (1/5000 dilution) was used for Western blotting.
    Blocking Buffer and concentration: 5% NFDM/TBST.
    Diluting buffer and concentration: 5% NFDM/TBST.
    Observed band size: 54 kDa

  • Western blot - Anti-Fascin antibody [EP5902] (ab126772)
    Western blot - Anti-Fascin antibody [EP5902] (ab126772)
    All lanes : Anti-Fascin antibody [EP5902] (ab126772) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 whole cell lysate
    Lane 2 : FSCN1 (Fascin) knockout HAP1 whole cell lysate
    Lane 3 : HepG2 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Predicted band size: 54 kDa



    Lanes 1 - 3: Merged signal (red and green). Green - ab126772 observed at 54 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab126772 was shown to specifically react with Fascin in wild-type HAP1 cells as signal was lost in FSCN1 (Fascin) knockout cells. Wild-type and FSCN1 (Fascin) knockout samples were subjected to SDS-PAGE. Ab126772 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Fascin antibody [EP5902] (ab126772)
    Western blot - Anti-Fascin antibody [EP5902] (ab126772)
    All lanes : Anti-Fascin antibody [EP5902] (ab126772) at 1/10000 dilution

    Lane 1 : SH SY5Y cell lysate
    Lane 2 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit at 1/2000 dilution

    Predicted band size: 54 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)

    ab126772, at 1/250 dilution, staining Fascin in paraffin-embedded Human colon tissue by Immunohistochemistry.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)

    ab126772 showing positive staining in Normal tonsil tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)

    ab126772 showing positive staining in Glioma tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)

    ab126772 showing positive staining in Normal kidney tissue.

    Perform heat mediated antigen retrieval before commencing with IHC staining protocol.

References (14)

Publishing research using ab126772? Please let us know so that we can cite the reference in this datasheet.

ab126772 has been referenced in 14 publications.

  • Chetry M  et al. Effects of Galectin-1 on Biological Behavior in Cervical Cancer. J Cancer 11:1584-1595 (2020). PubMed: 32047564
  • Fan YL  et al. A function of fascin1 in the colony formation of mouse embryonic stem cells. Stem Cells N/A:N/A (2020). PubMed: 32379912
  • van de Willige D  et al. Cytolinker Gas2L1 regulates axon morphology through microtubule-modulated actin stabilization. EMBO Rep 20:e47732 (2019). PubMed: 31486213
  • Chen Y  et al. FSCN1 is an effective marker of poor prognosis and a potential therapeutic target in human tongue squamous cell carcinoma. Cell Death Dis 10:356 (2019). PubMed: 31043585
  • Shen SN  et al. Down-regulation of long noncoding RNA PVT1 inhibits esophageal carcinoma cell migration and invasion and promotes cell apoptosis via microRNA-145-mediated inhibition of FSCN1. Mol Oncol N/A:N/A (2019). PubMed: 31369196
View all Publications for this product

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