Overview

  • Product nameAnti-beta Actin antibody [AC-15]
    See all beta Actin primary antibodies
  • Description
    Mouse monoclonal [AC-15] to beta Actin
  • SpecificityThis antibody makes a good loading control antibody and we recommend its use as such. We also recommend ab8226, another excellent mouse monoclonal beta actin loading control antibody.
  • Tested applicationsICC/IF, Dot Blot, IHC-FoFr, ICC, IHC-P, IHC-Fr, Indirect ELISA, WB, ELISA, IHC-FrFlmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Sheep, Rabbit, Chicken, Guinea pig, Hamster, Cow, Cat, Dog, Human, Carp, Opossum, Marmoset (common), Meriones unguiculatus

    Does not react with

    Fruit fly (Drosophila melanogaster), Dictyostelium discoideum
  • Immunogen

    Synthetic peptide corresponding to beta Actin aa 1-14 (N terminal) conjugated to Keyhole Limpet Haemocyanin (KLH). Slightly modified β-cytoplasmic actin N-terminal peptide, Ac-Asp-Asp-Asp-Ile-Ala-Al​a-Leu-Val-Ile-Asp-Asn-Gl y​-Ser-Gly-Lys, conjugated to KLH.
    Sequence:

    DDDIAALVIDNGSGK

  • EpitopeN-terminal of the beta isoform of actin.
  • Positive control
    • Cultured human or chicken fibroblasts as described in Liao et al. ICC/IF: SV40 cells
  • General notes

    Immunofluoresence staining of chicken gizzard ultra-thin sections (North et al. J. Cell Sci. 107:445-455 (1994)) labels the dense bodies, longitudinal channels and membrane associated dense-plaque.

Properties

Applications

Our Abpromise guarantee covers the use of ab6276 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
ICC/IF 1/200.
Dot Blot Use at an assay dependent concentration.
IHC-FoFr Use at an assay dependent concentration.
ICC 1/1000.
IHC-P 1/10000. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
IHC-Fr Use at an assay dependent concentration.
Indirect ELISA Use at an assay dependent concentration.
WB 1/5000 - 1/16000. Predicted molecular weight: 42 kDa.
ELISA Use at an assay dependent concentration.
IHC-FrFl Use at an assay dependent concentration.
IF Use a concentration of 5 µg/ml.
  • Application notesIs unsuitable for IP.
  • 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 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 similaritiesBelongs to the actin family.
    • Post-translational
      modifications
      ISGylated.
    • Cellular localizationCytoplasm > cytoskeleton. Localized in cytoplasmic mRNP granules containing untranslated mRNAs.
    • Information by UniProt
    • Database links
    • Alternative names
      • A X actin like protein antibody
      • ACTB antibody
      • ACTB_HUMAN antibody
      • Actin antibody
      • Actin cytoplasmic 1 antibody
      • actin, beta antibody
      • Actx antibody
      • Beta cytoskeletal actin antibody
      • Beta-actin antibody
      • BRWS1 antibody
      • cytoplasmic 1 antibody
      • Melanoma X actin antibody
      • N-terminally processed antibody
      • PS1TP5 binding protein 1 antibody
      • PS1TP5-binding protein 1 antibody
      • PS1TP5BP1 antibody
      see all

    Anti-beta Actin antibody [AC-15] images

    • ab6276 staining beta Actin in SV40 cells. The cells were fixed with 4% formaldehyde (10 min), permeabilized in 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 ab6276 at a working concentration of 5μg/ml and ab190573, Rabbit monoclonal [EP1332Y] to alpha Tubulin (Alexa Fluor® 647, shown in red) at 1/250 overnight at +4°C, followed by a further incubation at room temperature for 1h with an anti-mouse AlexaFluor® 488 (ab150117) at 2 μg/ml (shown in green). Nuclear DNA was labelled in blue with DAPI.
      Image was taken with a confocal microscope (Leica-Microsystems, TCS SP8).



    • Predicted band size : 42 kDa

      David Grimm, Yale University, USA.

      MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard asssay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.

      MDCK cells induced with increasing amounts of doxycycline to control expression of the gene of interest. All cells were normalized for loading with an albumin protein standard asssay. Anti-beta actin (ab6276) was used at a concentration of 1:5000 in a milk blocking solution. B-actin blotting confirms the albumin assay in showing that an equal amount of lysate was loaded in each lane.

    • All lanes : Anti-beta Actin antibody [AC-15] (ab6276) at 1/5000 dilution

      Lane 1 : HeLa nuclear
      Lane 2 : HeLa whole cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : Jurkat cell lysate
      Lane 5 : HEK293 cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      Alexa Fluor anti mouse at 1/5000 dilution

      Performed under reducing conditions.

      Predicted band size : 42 kDa
      Observed band size : 42 kDa


    • Predicted band size : 42 kDa
      Observed band size : 42 kDa

      This image is courtesy of an Abreview submitted by Dr Mark Elliott

      Western Blot of ab6276 (used as loading control) with whole tissue lysate of grey matter from BA20 (temporal cortex).  Ab6276 was diluted 1/50000 and incubated with the sample for 16 hours at 4°C.  5 µg of lysate was loaded onto the gel, which was blocked with 5% milk for 1 hour at 20°C.  An Alexa Fluor® 680 conjugated goat anti-mouse antibody, diluted 1/5000, was used as the secondary.

      Bands below actin in image are synaptophysin (SYN).

      See Abreview

    • ab6276 at 1/500 staining human macrophage cells by ICC/IF. The cells were paraformaldehyde fixed and blocked with serum prior to incubation with the antibody for 1 hour. A Cy2 ® conjugated donkey antibody was used as the secondary.

      See Abreview

    • ab6276 at 1/10000 staining mouse brain tissue sections by IHC-P. The mouse brain was immersion-fixed in 4% Formalin/PBS for 2 days, then bisected mid-saggitally. The tissue was further fixed in same solution for a further 3 days. Both halves were processed to pwax on a dip-n-dunk tissue processor using a 20hr-cycle. Sections were cut at 5microns,floated out on water at 47C, mounted on Superfrost Plus slides and dried/melted for 24hrs in a 62C oven.

      The whole brain shows positivity but there are areas/cells that are enriched. Note that the axonal tracks are negatively stained at this dilution factor, yet some cells within those tracks are still strongly positive (many similar cells outside of the tracks are also positive).

      See Abreview

    • ab6276 staining beat actin in rat hypothalamus primary cells by ICC/IF. The cells were formaldehyde fixed, permeabilized in 0.5% Triton X-100 and blocked in 5% serum for 20 minutes at 25°C. The primary antibody was diluted 1/1000 in PBS (0.1% Triton X-100, 1% goat serum) and incubated with sample for 16 hours at 4°C. An Alexa Fluor® 546 conjugated goat polyclonal to mouse IgG1, diluted 1/500 was used as secondary.

      See Abreview

    • ab6276 detecting beta actin in AGS Human gastric carcinoma cells by direct ELISA. Samples were blocked with 5% BSA for 1 hour at 23°C and incubated with the primary antibody (1/1000 in blocking buffer) for 16 hours at 4°C. Ab6729 (1/1000) was used as the secondary antibody.

      See Abreview

    References for Anti-beta Actin antibody [AC-15] (ab6276)

    This product has been referenced in:
    • Piskareva O  et al. The development of cisplatin resistance in neuroblastoma is accompanied by epithelial to mesenchymal transition in vitro. Cancer Lett N/A:N/A (2015). Read more (PubMed: 25960282) »
    • Lundby A  et al. Surface-expressed insulin receptors as well as IGF-I receptors both contribute to the mitogenic effects of human insulin and its analogues. J Appl Toxicol 35:842-50 (2015). Read more (PubMed: 25413577) »

    See all 473 Publications for this product

    Product Wall

    Application Western blot
    Loading amount 30 µg
    Gel Running Conditions Reduced Denaturing (12)
    Sample Human Tissue lysate - whole (CNS tissue)
    Specification CNS tissue
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Submitted Aug 01 2014

    Application Immunocytochemistry/ Immunofluorescence
    Blocking step BSA as blocking agent for 45 minute(s) · Concentration: 2% · Temperature: RT°C
    Sample Human Cell (HeLa cells)
    Specification HeLa cells
    Permeabilization Yes - 0.2% Triton-X100
    Fixative Paraformaldehyde
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    Submitted May 23 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 25 µg
    Gel Running Conditions Reduced Denaturing (12.5%)
    Sample Mouse Cell lysate - whole cell (Mouse brain lysates)
    Specification Mouse brain lysates
    Blocking step Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Herr Dr. Vladimir Milenkovic

    Verified customer

    Submitted Jul 10 2013

    Application Western blot
    Sample Rat Cell lysate - whole cell (h9c2)
    Gel Running Conditions Reduced Denaturing
    Loading amount 30 µg
    Specification h9c2
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    Submitted Jun 26 2015

    Application Western blot
    Sample Chinese Hamster Cell lysate - whole cell (CHO cell)
    Gel Running Conditions Reduced Denaturing
    Loading amount 30 µg
    Specification CHO cell
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    Submitted Jun 26 2015

    Application Western blot
    Sample Mouse Cell lysate - whole cell (Mouse inner medullary collecting duct (IMCD3) cell)
    Gel Running Conditions Reduced Denaturing
    Loading amount 20 µg
    Specification Mouse inner medullary collecting duct (IMCD3) cell
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Submitted Jun 23 2015

    Application Western blot
    Sample Human Cell lysate - whole cell (Fibroblast)
    Gel Running Conditions Reduced Denaturing
    Loading amount 20 µg
    Specification Fibroblast
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
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    Submitted Jun 23 2015

    Application Western blot
    Sample Human cytomegalovirus Cell lysate - whole cell (HCMV-infected human foreskin fibroblasts)
    Gel Running Conditions Reduced Denaturing (12.5% gel)
    Loading amount 500000 cells
    Treatment 1 µM Shield-1
    Specification HCMV-infected human foreskin fibroblasts
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Dr. Yi-Chieh Perng

    Verified customer

    Submitted Jun 22 2015

    Application Western blot
    Sample Human Cell lysate - whole cell (HEK293)
    Gel Running Conditions Reduced Denaturing
    Loading amount 30 µg
    Specification HEK293
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    Submitted Jun 22 2015

    Application Western blot
    Sample Mouse Cell lysate - whole cell (MEF)
    Gel Running Conditions Reduced Denaturing
    Loading amount 30 µg
    Specification MEF
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
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    Submitted Jun 22 2015

    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"