Product nameAnti-Actin antibody [C4]
See all Actin primary antibodies
DescriptionMouse monoclonal [C4] to Actin
Specificityab14128 recognizes all six isoforms of vertebrate actin, globular (G) and filamentous (F) forms; it is a pan-actin antibody that binds to an epitope in a highly conserved region of actin. ab14128 labels myotubes and stains myoblasts and fibroblasts, it does not interfere with actin polymerization to form filaments, at a ratio as high as one antibody per two actin monomers. In ELISA, ab14128 shows a strong reactivity with cytoplasmic actin and significant binding to gizzard, skeletal, arterial and cardiac actins, the antibody also shows significant binding to both Dictyostelium discoidum and Physarum polycephalum in ELISA. The clone number has been updated from (2Q1055) to (C4) both clone numbers name the same antibody clone.
Tested applicationsSuitable for: ICC/IF, WB, ICCmore details
Species reactivityReacts with: Chicken, Saccharomyces cerevisiae, Tetrahymena, Caenorhabditis elegans, Mammals, Dictyostelium discoideum, Physarum polycephalum, Vertebrata
Recombinant fragment corresponding to Chicken Actin aa 50-70 (N terminal). The immuogen is purified chicken gizzard actin. This is predominantly smooth muscle gamma-actin. Is a pan-actin which recognizes all actin isoforms.
Database link: P63270
EpitopeThe epitope recognized by the antibody appears to be located in the N-terminal two thirds of the actin molecule, possibly near amino acids 50-70.
- This antibody gave a positive signal in both Human Skeletal Muscle and Heart tissue lysates as well as the following whole cell lysates: A431; HEK293; MCF7; HeLa; MDA MB 231. ICC/IF: HeLa cells
General notesFor maximum recovery of product, centrifuge the original vial after thawing and prior to removing the cap. Further dilutions can be made in assay buffer.
Storage instructionsShipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C. Avoid freeze / thaw cycle.
Storage bufferpH: 7.40
Preservative: 0.05% Sodium azide
Constituents: Tris glycine, 0.88% Sodium chloride
Concentration information loading...
PurityProtein G purified
Purification notesPurified by Protein G affinity chromatography.
Light chain typekappa
Our Abpromise guarantee covers the use of ab14128 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
|ICC/IF||Use a concentration of 5 µg/ml.|
|WB||Use a concentration of 1 - 20 µg/ml. Predicted molecular weight: 43 kDa.
On muscle homogenates subject to SDS-PAGE, reacts relatively uniformly with a 43kDa protein present in skeletal, cardiac, gizzard and aorta tissues. Appears to react with all isoforms of actin found in these preparations and shows a strong reaction with the alpha actin found in skeletal, cardiac and arterial muscle.
Methanol fixed HeLa and NIH/3T3 cells
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
- a actin antibody
- ACTA antibody
- ACTA1 antibody
All lanes : Anti-Actin antibody [C4] (ab14128) at 1/1000 dilution
Lane 1 : A431 (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 2 : HEK293 (Human embryonic kidney cell line) Whole Cell Lysate
Lane 3 : MCF7 (Human breast adenocarcinoma cell line) Whole Cell Lysate
Lane 4 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
Lane 5 : MDA-MB-231 (Breast Carcinoma cell line)
Lane 6 : Skeletal Muscle (Human) Tissue Lysate - adult normal tissue
Lane 7 : Heart (Human) Tissue Lysate - adult normal tissue
Lysates/proteins at 20 µg per lane.
All lanes : Goat Anti-Mouse IgG H&L (Alexa Fluor® 790) (ab175783) at 1/10000 dilution
Predicted band size: 43 kDa
Observed band size: 42 kDa why is the actual band size different from the predicted?
This blot was produced using a 4-12% Bis-tris gel under the MOPS buffer system. The gel was run at 200V for 50 minutes before being transferred onto a Nitrocellulose membrane at 30V for 70 minutes. The membrane was then blocked for an hour using 5% Milk before being incubated with ab14128 overnight at 4°C. Antibody binding was detected using ab175783 at a 1:10,000 dilution for 1hr at room temperature and then imaged using the Licor Odyssey CLx.
ab14128 staining Actin in HeLa cells. The cells were fixed with 100% methanol (5min), permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked in 1% BSA/10% normal goat serum/0.3M glycine in 0.1%PBS-Tween for 1h. The cells were then incubated with ab14128 at 5μg/ml and ab6046 at 1µg/ml overnight at +4°C, followed by a further incubation at room temperature for 1h with an Goat anti-Mouse Alexa Fluor 488 secondary (ab150117) at 2 μg/ml (shown in green) and Goat anti-Rabbit Alexa Fluor 594 secondary (ab150084) at 2 μg/ml (shown in pseudo color red). Nuclear DNA was labelled in blue with DAPI.
Negative controls: 1– Rabbit primary and anti-mouse secondary antibody; 2 – Mouse primary antibody and anti-rabbit secondary antibody. Controls 1 and 2 indicate that there is no unspecific reaction between primary and secondary antibodies used.
This product has been referenced in:
- Wischhof L et al. A disease-associated Aifm1 variant induces severe myopathy in knockin mice. Mol Metab N/A:N/A (2018). Read more (PubMed: 29780003) »
- Leite-Dellova DCA et al. Signaling pathways involved in the rapid biphasic effect of aldosterone on Na+/H+ exchanger in rat proximal tubule cells. J Steroid Biochem Mol Biol N/A:N/A (2018). Read more (PubMed: 29702262) »