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  1. Link

    fascin-antibody-ep5902-ab126772.pdf

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Signal Transduction Cytoskeleton / ECM Cytoskeleton Microfilaments Actin etc Actin Binding Proteins
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Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Fascin antibody [EP5902] (ab126772)

  • Datasheet
  • SDS
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Western blot - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunocytochemistry/ Immunofluorescence - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
  • Flow Cytometry (Intracellular) - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunoprecipitation - Anti-Fascin antibody [EP5902] (ab126772)
  • Western blot - Anti-Fascin antibody [EP5902] (ab126772)
  • Western blot - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] (ab126772)
  • Anti-Fascin antibody [EP5902] (ab126772)

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: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

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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: Flow Cyt (Intra), WB, IP, IHC-P, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Human
    Predicted to work with: Rat
  • 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 (intra): 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 (glycerin, glycerine), 0.05% BSA, 50% Tissue culture supernatant
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP5902
  • Isotype

    IgG
  • Research areas

    • Signal Transduction
    • Cytoskeleton / ECM
    • Cytoskeleton
    • Microfilaments
    • Actin etc
    • Actin Binding Proteins

Associated products

  • Alternative Versions

    • Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
  • Compatible Secondaries

    • Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077)
    • Goat Anti-Rabbit IgG H&L (HRP) (ab205718)
    • Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773)
  • Isotype control

    • Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
  • KO cell lines

    • Human FSCN1 (Fascin) knockout HCT116 cell line (ab266895)
  • KO cell lysates

    • Human FSCN1 (Fascin) knockout HCT116 cell lysate (ab257444)
  • KO cell pellets

    • Human FSCN1 (Fascin) knockout HCT116 cell pellet (ab278993)
  • Recombinant Protein

    • Recombinant Human Fascin protein (ab99240)

Applications

The Abpromise guarantee

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
Flow Cyt (Intra)
1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.)
ICC/IF
1/500.

For unpurified use at 1/100 - 1/250

Notes
Flow Cyt (Intra)
1/100 - 1/1000.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

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.)
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.
  • Target information above from: UniProt accession Q16658 The UniProt Consortium
    The Universal Protein Resource (UniProt) in 2010
    Nucleic Acids Res. 38:D142-D148 (2010) .

    Information by UniProt
  • Database links

    • Entrez Gene: 6624 Human
    • Entrez Gene: 14086 Mouse
    • Entrez Gene: 683788 Rat
    • Omim: 602689 Human
    • SwissProt: Q16658 Human
    • SwissProt: Q61553 Mouse
    • SwissProt: P85845 Rat
    • Unigene: 118400 Human
    • Unigene: 289707 Mouse
    • Unigene: 199526 Rat
    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 (Intracellular) - Anti-Fascin antibody [EP5902] (ab126772)
    Flow Cytometry (Intracellular) - 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.

  • Anti-Fascin antibody [EP5902] (ab126772)
    Anti-Fascin antibody [EP5902] (ab126772)

Protocols

  • Flow cytometry protocols
  • Immunoprecipitation protocols
  • Immunohistochemistry protocols
  • Immunocytochemistry & immunofluorescence protocols
  • Western blot protocols

Click here to view the general protocols

Datasheets and documents

  • SDS download

  • Datasheet download

    Download

References (25)

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

ab126772 has been referenced in 25 publications.

  • Moroki T  et al. Databases for technical aspects of immunohistochemistry: 2021 update. J Toxicol Pathol 34:161-180 (2021). PubMed: 33976473
  • Xie F  et al. CircPTPRA blocks the recognition of RNA N6-methyladenosine through interacting with IGF2BP1 to suppress bladder cancer progression. Mol Cancer 20:68 (2021). PubMed: 33853613
  • Tang J  et al. LncRNA LOC146880 promotes esophageal squamous cell carcinoma progression via miR-328-5p/FSCN1/MAPK axis. Aging (Albany NY) 13:14198-14218 (2021). PubMed: 34016787
  • Goveia J  et al. An Integrated Gene Expression Landscape Profiling Approach to Identify Lung Tumor Endothelial Cell Heterogeneity and Angiogenic Candidates. Cancer Cell 37:21-36.e13 (2020). PubMed: 31935371
  • Lionetti MC  et al. Chromatin and Cytoskeletal Tethering Determine Nuclear Morphology in Progerin-Expressing Cells. Biophys J 118:2319-2332 (2020). PubMed: 32320674
View all Publications for this product

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