Overview

  • Product nameAnti-Actin antibody [ACTN05 (C4)]
    See all Actin primary antibodies
  • Description
    Mouse monoclonal [ACTN05 (C4)] to Actin
  • SpecificityThis antibody reacts with all six known vertebrate isoforms of actin. It also reacts with two cytoplasmic actins (beta, lambda) which are highly homologous to one another but which differ from the muscle actins at about 25 amino acidresidues.
  • Tested applicationsFlow Cyt, IP, WB, IHC-Fr, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Rabbit, Cow, Dog, Human, Pig, Dictyostelium discoideum, Physarum polycephalum
    Predicted to work with: Chicken
  • Immunogen

    Chicken gizzard actin.

  • Positive control
    • HeLa cells.

Properties

Applications

Our Abpromise guarantee covers the use of ab3280 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 1-2µg for 106 cells.
IP Use at 2 µg/mg of lysate. Use Protein G. This has been tested in IP on "denatured" cell lysate (treated with 5% SDS).
WB Use a concentration of 0.5 µg/ml. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IHC-Fr 1/250.
IHC-P Use a concentration of 0.5 - 1 µg/ml. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent dilution. See Bryce et al. Fix cells with 4% paraformaldehyde and permeabilize with chilled methanol.

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
    • Beta cytoskeletal actin 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 (C4)] images

  • All lanes : Anti-Actin antibody [ACTN05 (C4)] (ab3280)

    Lane 1 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 2 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate
    Lane 3 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 4 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 5 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate
    Lane 6 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
    Lane 7 : NIH 3T3 (Mouse embryonic fibroblast cell line) Whole Cell Lysate
    Lane 8 : MDA-MB-231 (Human breast adenocarcinoma cell line) Whole Cell Lysate Whole Cell Lysate
    Lane 9 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate

    Lysates/proteins at 10 µg per lane.

    developed using the ECL technique

    Performed under reducing conditions.

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

    Lanes: 1-3: 4oC (x1 freeze/thaw)

    Lanes: 4-6: 4oC (x2 freeze/thaws)

    Lanes 7-9:  4oC (x4 freeze/thaws)   

  • All lanes : Anti-Actin antibody [ACTN05 (C4)] (ab3280) at 1/2000 dilution

    Lane 1 : Mouse liver whole tissue lysate (mouse 1).
    Lane 2 : Mouse liver whole tissue lysate (mouse 2).
    Lane 3 : Mouse liver whole tissue lysate (mouse 3).

    Lysates/proteins at 50 µg per lane.

    Secondary
    An HRP-conjugated goat anti-mouse IgG polyclonal. at 1/5000 dilution
    developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 42 kDa

    This image is courtesy of an anonymous Abreview

    Blocking Step: 5% Milk for 30 minutes at 25°C.

    See Abreview

  • ab3280 staining Actin in mouse heart tissue sections by IHC-Fr (formaldehyde-fixed frozen sections). Tissue samples were fixed with formaldehyde, permeabilized with 0.1% Triton X-100 and blocked with 5% serum for 1 hour at 25°C. The sample was incubated with primary antibody (1/250 in PBS-Tween) at 4°C for 12 hours. An Alexa Fluor®488-conjugated Goat polyclonal to mouse IgG (1/250) was used as secondary antibody. Nuclei were stained with DAPI.

    See Abreview

  • ab3280 (1µg/ml) staining actin in human colon using an automated system (DAKO Autostainer Plus). Using this protocol there is strong cytoplasmic staining throughout the tissue.
    Sections were rehydrated and antigen retrieved with the Dako 3 in 1 AR buffer citrate pH6.1 in a DAKO PT link. Slides were peroxidase blocked in 3% H2O2 in methanol for 10 mins. They were then blocked with Dako Protein block for 10 minutes (containing casein 0.25% in PBS) then incubated with primary antibody for 20 min and detected with Dako envision flex amplification kit for 30 minutes. Colorimetric detection was completed with Diaminobenzidine for 5 minutes. Slides were counterstained with Haematoxylin and coverslipped under DePeX. Please note that, for manual staining, optimization of primary antibody concentration and incubation time is recommended. Signal amplification may be required.

  • ICC/IF image of ab3280 stained Hepp cells. The cells were 4% formaldehyde fixed (10 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 (ab3280, 10µg/ml) overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti-mouse IgG (H+L) used at a 1/1000 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.

  • ICC/IF image of ab3280 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 (ab3280, 5µg/ml) overnight at +4°C. The secondary antibody (green) was DyLight® 488 goat anti-mouse IgG - H&L, pre-adsorbed (ab96879) 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.
  • Overlay histogram showing HeLa cells stained with ab3280 (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 (ab3280, 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 (C4)] (ab3280)

This product has been referenced in:
  • Yim DG  et al. Casein kinase 1 regulates Sprouty2 in FGF-ERK signaling. Oncogene N/A:N/A (2014). Read more (PubMed: 24469046) »
  • Balzamino BO  et al. Characterization of NGF, trkA (NGFR) , and p75 (NTR) in Retina of Mice Lacking Reelin Glycoprotein. Int J Cell Biol 2014:725928 (2014). WB ; Mouse . Read more (PubMed: 24627687) »

See all 60 Publications for this product

Product Wall

Application Western blot
Loading amount 20 µg
Gel Running Conditions Reduced Denaturing (10 % GEL, SDS)
Sample Human Cell lysate - whole cell (HEK293)
Specification HEK293
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Apr 23 2014

Thank you for contacting us and your interest in our products.

As discussed over the phone, the anti-actin antibody ab3280 has been shown to reacts with all six known vertebrate isoforms of actin. It also reacts with two cytoplasmic actins (be...

Read More

Thank you for your reply.
We have beta actin antibodies that would fulfill all your criteria:
ab8227 (http://www.abcam.com/beta-Actin-antibody-Loading-Control-ab8227.html) - in case beta actin would be acceptable for your experiments.
Or...

Read More

Thank you for your reply.
If the word "actin" is entered into the search box on our homepage (top right corner), you will obtain entries for 21 actin antibodies, 9 actin-loading control antibodies, etc.
We do not have a rabbit anti-actin anti...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (Hepatoma)
Loading amount 25 µg
Specification Hepatoma
Gel Running Conditions Reduced Denaturing (10)
Blocking step Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 4°C
Username

Mrs. Alina Macovei

Verified customer

Submitted May 21 2012

Thank you for contacting us.

Anti actin or anti GAPDH antibodies are most suitable to be used as loading control.

Please recommend customer ab3280 or ab9485.

I hope this information is helpful to you. Please do not hesit...

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Thank you for your recent telephone enquiries.

I have listed below some primary antibodies that may be of interest to you. I can recommend to review the online datasheets for further information.

Actin
I am sorry there are no...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK293)
Loading amount 20 µg
Specification HEK293
Treatment Lane 1-No treatment, lane 2-30uM DMSO overnight, lane 3-20uM DMSO overnight
Gel Running Conditions Non-reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 21°C
Username

Abcam user community

Verified customer

Submitted Jan 30 2012

Thank you for contacting us.

I am sorry that this antibody did not perform as stated on the datasheet. If payment has already been made on the original order and you wish to receive a refund, please ask your purchasing department to contact ...

Read More
Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Sample Human Cell lysate - whole cell (HEK 293 cells)
Loading amount 100 µg
Specification HEK 293 cells
Gel Running Conditions Reduced Denaturing (10% gel)
Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Abcam user community

Verified customer

Submitted Jan 16 2012

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