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

Mouse monoclonal F-actin antibody [4E3.adl]. Validated in WB, Flow Cyt, ICC/IF and tested in Mouse, Human. Cited in 5 publication(s).

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

    Suitable for: WB, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    corresponding to F-actin.

  • 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab130935 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 1/500. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).

Abcam recommends blocking with 3% Milk

ICC/IF Use a concentration of 5 µg/ml.
Flow Cyt Use 0.1-1µg for 106 cells.

ab91545 - Mouse monoclonal IgM, is suitable for use as an isotype control with this antibody.

Target

  • Function

    Actins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • Involvement in disease

    Defects in ACTB are a cause of dystonia juvenile-onset (DYTJ) [MIM:607371]. DYTJ is a form of dystonia with juvenile onset. Dystonia is defined by the presence of sustained involuntary muscle contraction, often leading to abnormal postures. DYTJ patients manifest progressive, generalized, dopa-unresponsive dystonia, developmental malformations and sensory hearing loss.
  • Sequence similarities

    Belongs to the actin family.
  • Post-translational
    modifications

    ISGylated.
  • Cellular localization

    Cytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
  • Information by UniProt
  • Database links

  • Alternative names

    • ACTB antibody
    • ACTB_HUMAN antibody
    • Actin, cytoplasmic 1, N-terminally processed antibody
    • Beta-actin antibody
    • f actin antibody
    • Filamentous actin antibody
    see all

Images

  • 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
  • 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.

  • 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.

References

This product has been referenced in:

  • Liu Y  et al. Reversal of TET-mediated 5-hmC loss in hypoxic fibroblasts by ascorbic acid. Lab Invest N/A:N/A (2019). Read more (PubMed: 30837678) »
  • Zeng S  et al. Transcriptome sequencing identifies ANLN as a promising prognostic biomarker in bladder urothelial carcinoma. Sci Rep 7:3151 (2017). IHC-P, IF ; Human . Read more (PubMed: 28600503) »
See all 6 Publications for this product

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