Astana Biomed Group, an authorized Abcam distributor in Central Asia

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

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 - BSA and Azide free
  • Suitable for: IHC-P, IP, ICC/IF, WB, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-Fascin antibody [EP5902] - BSA and Azide free
    See all Fascin primary antibodies

  • Description

    Rabbit monoclonal [EP5902] to Fascin - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    IP
    Human
    WB
    Human
    See all applications and species data

  • Immunogen

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

  • Positive control

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

  • General notes

    Ab240034 is the carrier-free version of ab126772. This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    ab240034 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

    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 the problem with our range of recombinant monoclonal antibodies and knockout edited cell lines for gold-standard validation.

    One factor contributing to the crisis is the use of antibodies that are not suitable. This can lead to misleading results and the use of incorrect data informing project assumptions and direction. To help address this challenge, we have introduced an application and species grid on our primary antibody datasheets to make it easy to simplify identification of the right antibody for your needs.

    Learn more here.

Properties

Images

  • Western blot - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Western blot - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab126772).

    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] - BSA and Azide free (ab240034)
    Immunocytochemistry/ Immunofluorescence - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)

    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. 

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

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

  • Flow Cytometry - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Flow Cytometry - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

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

    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
    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)

    ab126772 showing positive staining in Normal tonsil tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)

    ab126772 showing positive staining in Glioma tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

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

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Fascin antibody [EP5902] - BSA and Azide free (ab240034)

    ab126772 showing positive staining in Normal kidney tissue.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126772).

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

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

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