Overview

  • Product name
    Anti-Actin antibody [EPR16769]
    See all Actin primary antibodies
  • Description
    Rabbit monoclonal [EPR16769] to Actin
  • Host species
    Rabbit
  • Tested applications
    Suitable for: WB, IHC-P, ICC/IF, IP, Flow Cytmore details
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Human
  • Immunogen

    Synthetic peptide within Human Actin aa 50-150. The exact sequence is proprietary.
    Database link: P68133

  • Positive control
    • WB: HeLa, 293T, C6, RAW 264.7, PC-12, NIH/3T3 and UMNSAH/DF-1 whole cell lysates; Human skeletal muscle, fetal spleen, fetal brain, fetal heart, fetal kidney and cardiac muscle lysates; Mouse and Rat brain, heart, kidney and spleen lysates. IHC-P: Human prostate hyperplasia, Mouse skeletal muscle and Rat skeletal muscle tissues. ICC/IF: NIH/3T3 cells. IP: NIH/3T3 whole cell extract. FC: HeLa cells
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Applications

Our Abpromise guarantee covers the use of ab179467 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 1/5000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa).
IHC-P 1/500 - 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/50.
IP 1/40.
Flow Cyt 1/70.

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 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 similarities
    Belongs to the actin family.
  • Cellular localization
    Cytoplasm > cytoskeleton.
  • Information by UniProt
  • Database links
  • Alternative names
    • a actin antibody
    • 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

Images

  • All lanes : Anti-Actin antibody [EPR16769] (ab179467) at 1/20000 dilution

    Lane 1 : Mouse brain lysate
    Lane 2 : Mouse heart lysate
    Lane 3 : Mouse kidney lysate
    Lane 4 : Mouse spleen lysate
    Lane 5 : Rat brain lysate
    Lane 6 : Rat heart lysate
    Lane 7 : Rat kidney lysate
    Lane 8 : Rat spleen lysate
    Lane 9 : C6 (Rat glial tumor cells) whole cell lysates
    Lane 10 : RAW 264.7 (Mouse macrophage cells transformed with Abelson murine leukemia virus) whole cell lysate
    Lane 11 : PC-12 (Rat adrenal gland pheochromocytoma) whole cell lysate
    Lane 12 : NIH/3T3 (Mouse embyro fibroblast cells) whole cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Human prostate hyperplasia tissue labeling Actin with ab179467 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm staining on smooth muscle cells is observed. Counter stained with Hematoxylin.


    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Immunocytochemistry/ Immunofluorescence analysis of NIH/3T3 (Mouse embryonic fibroblast) cells labeling Actin with Purified ab179467 at 1:100 dilution (6.98 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 dilution (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunocytochemistry/ Immunofluorescence analysis of HeLa (Human cervix adenocarcinoma epithelia cell) cells labeling Actin with Purified ab179467 at 1:100 dilution ( 6.98 µg/ml). Cells were fixed in 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. Cells were counterstained with Ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) 1:200 (2.5 µg/ml). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077) was used as the secondary antibody at 1:1000 dilution (2 µg/ml) dilution. DAPI nuclear counterstain. PBS instead of the primary antibody was used as the secondary antibody only control.

  • Immunohistochemical analysis of paraffin-embedded Mouse skeletal muscle tissue labeling Actin with ab179467 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm staining is observed. Counter stained with Hematoxylin.


    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized NIH/3T3 (Mouse embyro fibroblast cells) cells labeling Actin with ab179467 at 1/50 dilution, followed by Goat anti-rabbit Alexa Fluor® 488 (IgG) (ab150077) secondary antibody at 1/200 dilution (green). Cytoplasm staining on NIH/3T3 cell line is observed. The nuclear counter stain is DAPI (blue). Tubulin is detected with ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution and ab150120 (goat anti-mouse AlexaFluor®594 secondary) at 1/500 dilution (red).
    The negative controls are as follows;
    1. ab179467 at 1/50 dilution followed by ab150120 (goat anti-mouse AlexaFluor®594 secondary) at 1/500 dilution.
    2. ab7291 (anti-Tubulin mouse mAb) at 1/500 dilution followed by ab150077 (goat anti-rabbit Alexa Fluor®488 (IgG H&L) at 1/200 dilution.

  • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labelling Actin with purified ab179467 at 1/70 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. An Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/2000) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

  • All lanes : Anti-Actin antibody [EPR16769] (ab179467) at 1/20000 dilution

    Lane 1 : HeLa (Human epithelial cells from cervix adenocarcinoma) whole cell lysates
    Lane 2 : 293T (Human epithelial cells from embryonic kidney) whole cell lysates
    Lane 3 : Human skeletal muscle lysate
    Lane 4 : Human fetal spleen lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L),Peroxidase conjugated at 1/1000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Actin antibody [EPR16769] (ab179467) at 1/5000 dilution

    Lane 1 : Human fetal brain lysate
    Lane 2 : Human fetal heart lysate
    Lane 3 : Human fetal kidney lysate
    Lane 4 : Human fetal spleen lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • All lanes : Anti-Actin antibody [EPR16769] (ab179467) at 1/20000 dilution

    Lane 1 : UMNSAH/DF-1 (Transformed chicken embyronic fibroblast cells) whole cell lysates)
    Lane 2 : Human cardiac muscle lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated at 1/1000 dilution

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



    Blocking/Dilution buffer: 5% NFDM/TBST.

  • Immunohistochemical analysis of paraffin-embedded Rat skeletal muscle tissue labeling Actin with ab179467 at 1/1000 dilution, followed by prediluted HRP Polymer for Rabbit/Mouse IgG. Cytoplasm staining is observed. Counter stained with Hematoxylin.

     

    Negative control: Using PBS instead of primary ab, secondary ab is prediluted HRP Polymer for Rabbit/Mouse IgG.

  • Actin was immunoprecipitated from 1mg of NIH/3T3 (Mouse embyro fibroblast cells) whole cell extract with ab179467 at 1/40 dilution. Western blot was performed from the immunoprecipitate using ab179467 at 1/1000 dilution. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as secondary antibody at 1/1000 dilution. Lane 1: NIH/3T3 whole cell extract. Lane 2: PBS instead of NIH/3T3 whole cell extract.


    Blocking and dilution buffer and concentration: 5% NFDM/TBST.

References

This product has been referenced in:
  • Zhu X  et al. BMS-345541 inhibits airway inflammation and epithelial-mesenchymal transition in airway remodeling of asthmatic mice. Int J Mol Med 42:1998-2008 (2018). Read more (PubMed: 30015827) »
  • He J & Zhu J Collapsin Response Mediator Protein-2 Ameliorates Sevoflurane-Mediated Neurocyte Injury by Targeting PI3K-mTOR-S6K Pathway. Med Sci Monit 24:4982-4991 (2018). Read more (PubMed: 30018280) »
See all 14 Publications for this product

Customer reviews and Q&As

Application
Western blot
Sample
Saccharomyces cerevisiae Cell lysate - whole cell (S. cerevisiae)
Gel Running Conditions
Reduced Denaturing (12%)
Loading amount
20 µg
Treatment
1. untreated, 2. 10 mM D,L-Hcy for 0.5 hrs, 3. 10 mM L-Met for 0.5 hrs, 4. 10 mM L-Leu for 0.5 hrs, 5. untreated, 6. 10 mM D,L-Hcy for 4 hrs, 7. 10 mM L-Met for 4 hrs, 8. 10 mM L-Leu for 4 hrs
Specification
S. cerevisiae
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 4°C

Abcam user community

Verified customer

Submitted Feb 26 2018

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

Sign up