Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-Filamin A antibody [EP2405Y] (ab76289)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2405Y] to Filamin A
  • Suitable for: ICC/IF, WB, IHC-P, Flow Cyt
  • Knockout validated
  • Reacts with: Mouse, Rat, Human, African green monkey

Overview

  • Product name

    Anti-Filamin A antibody [EP2405Y]
    See all Filamin A primary antibodies

  • Description

    Rabbit monoclonal [EP2405Y] to Filamin A

  • Host species

    Rabbit

  • Tested Applications & Species

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

  • Immunogen

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

  • Positive control

    • COS-1, HeLa, HepG2, 3T3 and C6 cell lysate and human uterus tissue.

  • 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.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

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.

  • Storage buffer

    pH: 7.20
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EP2405Y

  • Isotype

    IgG

  • Research areas

Images

  • Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)

    Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: FLNA knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HepG2 whole cell lysate (20 µg)
    Lane 4: HeLa whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab76289 observed at 281 kDa. Red - loading control, ab18058, observed at 130 kDa.

    ab76289 was shown to specifically react with FLNA when FLNA knockout samples were used. Wild-type and FLNA knockout samples were subjected to SDS-PAGE. Ab76289 and ab18058 (Mouse anti Vinculin loading control) were incubated overnight at 4°C at 250000 dilution and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
    All lanes : Anti-Filamin A antibody [EP2405Y] (ab76289) at 1/10000 dilution

    Lane 1 : HeLa (human cervix adenocarcinoma) whole cell lysate
    Lane 2 : COS-1 (Cercopithecus aethiops kidney) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), HRP conjugated at 1/1000 dilution

    Predicted band size: 281 kDa
    Additional bands at: 281 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 30 seconds


    Blocking buffer: 5% NFDM/TBST

    Diluting buffer: 5% NFDM/TBST

  • Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)

    ab76289 staining Filamin A in the human cell line HeLa (human cervix adenocarcinoma) by flow cytometry. Cells were fixed with 4% paraformaldehyde, permeabilized with 90% methanol and the sample was incubated with the primary antibody at a dilution of 1/20. A goat anti rabbit IgG (Alexa Fluor® 488) at a dilution of 1/2000 was used as the secondary antibody.

    Isoytype control: Rabbit monoclonal IgG (Black)

    Unlabelled control: Cell without incubation with primary antibody and secondary antibody (Blue)

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Filamin A antibody [EP2405Y] (ab76289)

    Immunohistochemical analysis of paraffin-embedded human uterus using ab76289 at a 1/100 dilution.

    Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.

  • Immunocytochemistry/ Immunofluorescence - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Immunocytochemistry/ Immunofluorescence - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Immunofluorescent staining of HeLa cells using ab76289 at a 1/100 dilution.
  • Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Flow Cytometry - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Overlay histogram showing A431 cells stained with ab76289 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton 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 (ab76289 , 1/100 dilutio) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in A431 cells fixed with 4% paraformaldehyde/permeabilized in 0.1% PBS-Triton used under the same conditions.

  • Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
    Western blot - Anti-Filamin A antibody [EP2405Y] (ab76289)
    All lanes : Anti-Filamin A antibody [EP2405Y] (ab76289) at 1/500000 dilution

    Lane 1 : COS-1 (African green monkey kidney fibroblast-like cell line) cell lysate
    Lane 2 : Hela cell lysate
    Lane 3 : 3T3 cell lysate
    Lane 4 : C6 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP labelled goat anti-rabbit at 1/1000 dilution

    Predicted band size: 281 kDa
    Observed band size: 281 kDa

  • Anti-Filamin A antibody [EP2405Y] (ab76289)
    Anti-Filamin A antibody [EP2405Y] (ab76289)

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

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