Anti-Actin antibody [ACTN05; same as C4] (ab76548)

Overview

  • Product nameAnti-Actin antibody [ACTN05; same as C4]
    See all Actin primary antibodies
  • Description
    Mouse monoclonal [ACTN05; same as C4] to Actin
  • SpecificityThis antibody reacts with all six known vertebrate isoforms of actin. It also reacts with two cytoplasmic actins (beta and gamma).
  • Tested applicationsFlow Cyt, ICC/IF, WB, IP, IHC-Pmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Chicken, Cow, Dog, Human, Pig, Dictyostelium discoideum, Physarum polycephalum
  • Immunogen

    Chicken gizzard Actin

  • Positive control
    • This antibody gave a positive result when used in the following methanol fixed cell lines: HeLa. Skeletal muscle
  • General notes

    This antibody is highly recommended for monitoring total protein load on Western blots.

Properties

Applications

Our Abpromise guarantee covers the use of ab76548 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 Use 1µg for 106 cells.
ICC/IF Use a concentration of 10 µg/ml.
WB Use a concentration of 0.5 µg/ml. Predicted molecular weight: 42 kDa.
IP Use at 2 µg/mg of lysate. Use Protein G.
IHC-P 1/50. Boil tissue sections in 10mM citrate buffer, pH 6.0 for 10 - 20 minutes followed by cooling at room temperature for 20 minutes.

Target

  • FunctionActins are highly conserved proteins that are involved in various types of cell motility and are ubiquitously expressed in all eukaryotic cells.
  • Involvement in diseaseDefects in ACTA1 are the cause of nemaline myopathy type 3 (NEM3) [MIM:161800]. A form of nemaline myopathy. Nemaline myopathies are muscular disorders characterized by muscle weakness of varying severity and onset, and abnormal thread-or rod-like structures in muscle fibers on histologic examination. The phenotype at histological level is variable. Some patients present areas devoid of oxidative activity containg (cores) within myofibers. Core lesions are unstructured and poorly circumscribed.
    Defects in ACTA1 are a cause of myopathy congenital with excess of thin myofilaments (MPCETM) [MIM:161800]. A congenital muscular disorder characterized at histological level by areas of sarcoplasm devoid of normal myofibrils and mitochondria, and replaced with dense masses of thin filaments. Central cores, rods, ragged red fibers, and necrosis are absent.
    Defects in ACTA1 are a cause of congenital myopathy with fiber-type disproportion (CFTD) [MIM:255310]; also known as congenital fiber-type disproportion myopathy (CFTDM). CFTD is a genetically heterogeneous disorder in which there is relative hypotrophy of type 1 muscle fibers compared to type 2 fibers on skeletal muscle biopsy. However, these findings are not specific and can be found in many different myopathic and neuropathic conditions.
  • Sequence similaritiesBelongs to the actin family.
  • Cellular localizationCytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • ACTA antibody
    • ACTA1 antibody
    • Actin alpha skeletal muscle antibody
    • Actin antibody
    • actin, alpha 1, skeletal muscle 1 antibody
    • actin, alpha 1, skeletal muscle antibody
    • Actin, alpha skeletal muscle antibody
    • actina antibody
    • actine antibody
    • ACTS_HUMAN antibody
    • aktin antibody
    • alpha Actin 1 antibody
    • alpha skeletal muscle Actin antibody
    • alpha skeletal muscle antibody
    • alpha-actin antibody
    • Alpha-actin-1 antibody
    • ASMA antibody
    • CFTD antibody
    • CFTD1 antibody
    • CFTDM antibody
    • MPFD antibody
    • NEM1 antibody
    • NEM2 antibody
    • NEM3 antibody
    • nemaline myopathy type 3 antibody
    see all

Anti-Actin antibody [ACTN05; same as C4] images

  • ab76548 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 permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab76548 at 10µg/ml overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti- mouse (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. This antibody also gave a positive result in methanol fixed (100%, 5min) Hek293, HepG2, and MCF-7 cells at 10ug/ml.

  • Anti-Actin antibody [ACTN05; same as C4] (ab76548) at 0.5 µg/ml + Human cells 8

    Predicted band size : 42 kDa
    Observed band size : 42 kDa
  • Actin was immmunoprecipitated from human cells 8 using ab76548, followed by Western blotting using ab76548 at 0.5 µg/ml.
  • Overlay histogram showing HeLa cells stained with ab76548 (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 (ab76548, 1µg/1x106 cells) for 30 min at 22ºC. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22ºC. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a positive signal in HeLa cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween for 20 min used under the same conditions.

References for Anti-Actin antibody [ACTN05; same as C4] (ab76548)

ab76548 has not yet been referenced specifically in any publications.

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