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Anti-F-actin antibody [4E3.adl] (ab130935)

Key features and details

  • Mouse monoclonal [4E3.adl] to F-actin
  • Suitable for: WB, ICC/IF, Flow Cyt
  • Reacts with: Mouse, Human
  • Isotype: IgM

Overview

  • Product name

    Anti-F-actin antibody [4E3.adl]
    See all F-actin primary antibodies

  • Description

    Mouse monoclonal [4E3.adl] to F-actin

  • Host species

    Mouse

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    WB
    Mouse
    See all applications and species data

  • Immunogen

    corresponding to F-actin.
    Database link: P68139

  • Positive control

    • This antibody gave a positive signal in the following lysates: Human Skeletal Muscle Tissue; Mouse Skeletal Muscle Tissue; Mouse Heart Tissue; HeLa Whole Cell; C2C12 Whole Cell. This antibody gave a positive result in IF in the following Methanol fixed cell line: HeLa.

  • General notes

    This antibody clone is manufactured by Abcam.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

    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-F-actin antibody [4E3.adl] (ab130935)
    Western blot - Anti-F-actin antibody [4E3.adl] (ab130935)
    All lanes : Anti-F-actin antibody [4E3.adl] (ab130935) at 1/500 dilution

    Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 2 : Skeletal Muscle (Mouse) Tissue Lysate
    Lane 3 : Heart (Mouse) Tissue Lysate
    Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 5 : C2C12 (Mouse myoblast cell line) Whole Cell Lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Peroxidase- conjugated AffiniPure Goat Anti-mouse IgM (ab98112) at 1/3000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 42 kDa
    Observed band size: 42 kDa
    Additional bands at: 60 kDa, 70 kDa. We are unsure as to the identity of these extra bands.


    Exposure time: 10 seconds
  • Flow Cytometry - Anti-F-actin antibody [4E3.adl] (ab130935)
    Flow Cytometry - Anti-F-actin antibody [4E3.adl] (ab130935)

    Overlay histogram showing HeLa cells stained with ab130935 (red line). The cells were fixed with 4% paraformaldehyde (10 min) and then permeabilized with 0.1% PBS-Triton X-100 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 (ab130935, 1μg/1x106 cells) for 30 min at 22°C. The secondary antibody used was a goat anti-mouse  DyLight® 488 (IgM; mu chain) (ab97007) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgM [ICIGM] (ab91545, 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 antibody gave a positive signal in HeLa cells fixed with 80% methanol (5 min)/permeabilized with 0.1% PBS-Triton X-100 for 20 min used under the same conditions.

  • Immunocytochemistry/ Immunofluorescence - Anti-F-actin antibody [4E3.adl] (ab130935)
    Immunocytochemistry/ Immunofluorescence - Anti-F-actin antibody [4E3.adl] (ab130935)

    ab130935 stained HeLa cells. The cells were 100% methanol fixed (5 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab130935 at 5µg/ml overnight at +4°C. The secondary antibody (green) was a goat anti-mouse DyLight® 488 (ab96879) IgG (H+L) used at a 1/250 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1h. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43µM.

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