Overview

  • Product name
    Anti-Actin antibody - Loading Control
    See all Actin primary antibodies
  • Description
    Rabbit polyclonal to Actin - Loading Control
  • Host species
    Rabbit
  • Specificity
    This antibody recognises beta and gamma actin in Human samples. It probably also recognises all the other known forms of Human actin. This antibody detects a single clean band in Human, Mouse, Rat, Chicken and Drosophila samples. In Xenopus laevis a secondary band is detected at about 30kDa. We are unsure whether this is cross-reaction with another actin isoform or merely non-specific. In Cow a doublet is detected, which probably represents different forms of actin.
  • Tested applications
    Suitable for: WB, IHC-Pmore details
    Unsuitable for: ICC/IF
  • Species reactivity
    Reacts with: Mouse, Rat, Chicken, Cow, Human, Xenopus laevis, Drosophila melanogaster, Zebrafish
    Predicted to work with: Rabbit, Saccharomyces cerevisiae, Orangutan
  • Immunogen

    Synthetic peptide conjugated to KLH derived from within residues 350 to the C-terminus of Human Actin.

    Read Abcam's proprietary immunogen policy (Peptide available as ab13771.)

  • Positive control
    • HeLa whole cell lysate or mouse brain lysate. IHC-P - Human Colon FFPE tissue section

Applications

Our Abpromise guarantee covers the use of ab1801 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 1/1000. Detects a band of approximately 42 kDa (predicted molecular weight: 42 kDa). Block in 5% BSA. Blocking in milk significantly reduces the signal.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
  • Application notes
    Is unsuitable for ICC/IF.
  • 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

    • IHC image of ab1801 staining Actin in normal human colon formalin-fixed paraffin-embedded tissue sections*, performed on a Leica Bond. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab1801, 5μl/ml concentration, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX. No primary antibody was used in the negative control (shown on the inset).

      For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

      *Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre

    • All lanes : Anti-Actin antibody - Loading Control (ab1801) at 1 µg/ml

      Lane 1 : HeLa (Human epithelial carcinoma cell line) Whole Cell Lysate
      Lane 2 : Brain (Rat) Tissue Lysate
      Lane 3 : Brain (Mouse) Tissue Lysate
      Lane 4 : NIH 3T3 whole cell lysate (ab7179)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/50000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

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


      Exposure time: 1 minute


      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 2% Bovine Serum Albumin before being incubated with ab1801 overnight at 4°C. Antibody binding was detected using an anti-rabbit antibody conjugated to HRP, and visualised using ECL development solution ab133406

    • Western blot analysis of HeLa cells treated for 12 hours with hesperidin (h) (2.5 μg/ml, 4,01 μM), mangiferin (5 μg/ml, 11.84 μM) (m), and hesperidin (2.5 μg/ml, 4.01 μM) in a presence of mangiferin (5 μg/ml, 11.84 μM) (h+m). Immunoblotting was performed with the following primary antibodies: Bax (ab32503), BCL2 (ab59348), beta actin (ab1801), and caspase 8. After the washing steps, the membranes were incubated with goat anti-rabbit IgG (H+L) or with goat anti-mouse IgG (H+L) HRP-conjugated secondary antibodies and detected using ECL. Densitometry was performed using Image Lab software v. 4.1 (BioRad).

      Top panel: Following 12h of treatment of HeLa cells with hesperidin (h), mangiferin (m), and hesperidin in a presence of mangiferin (h+m), the mRNA levels were monitored in real - time PCR experiments. The BAX and BCL2 mRNA levels results from 2 independent experiments (n?=?8) are plotted relative to glyceraldehyde-3-phosphate dehydrogenase (GAPDH) and 18S rRNA levels and expressed as a fold change over the EtOH control. Error bars represent standard derivations.

      Bottom panel: Following 12h of treatment of HeLa cells with hesperidin (h), mangiferin (m), and hesperidin in a presence of mangiferin (h+m), the protein levels of Bax and BCL2 were detected with SDS-PAGE and Western Blot and related to beta actin levels.

    • All lanes : Anti-Actin antibody - Loading Control (ab1801) at 1 µg/ml

      Lane 1 : Brain (Mouse) Tissue Lysate (ab27253)
      Lane 2 : NIH 3T3 (Mouse) Whole Cell Lysate (ab52956)

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) preadsorbed (ab97080) at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 42 kDa


      Exposure time: 30 seconds
    • All lanes : Anti-Actin antibody - Loading Control (ab1801) at 1/1000 dilution

      All lanes : Whole cell lysates prepared from HUVEC cells

      Lysates/proteins at 15 µg per lane.

      Secondary
      All lanes : HRP-conjugated goat polyclonal to rabbit Ig at 1/5000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 42 kDa


      Exposure time: 30 seconds

      See Abreview

    References

    This product has been referenced in:
    • Kassahun H  et al. Constitutive MAP-kinase activation suppresses germline apoptosis in NTH-1 DNA glycosylase deficient C. elegans. DNA Repair (Amst) 61:46-55 (2018). Read more (PubMed: 29202295) »
    • Yang Y  et al. The novel dithiocarbamate, DpdtC suppresses HER2-overexpressed cancer cells by up-regulating NDRG1 via inactivation of HER2-ERK 1/2 signaling. Sci Rep 8:3398 (2018). Read more (PubMed: 29467385) »
    See all 121 Publications for this product

    Customer reviews and Q&As

    1-10 of 40 Abreviews or Q&A

    Application
    Western blot
    Sample
    Mouse Tissue lysate - whole (RIPA lysate of whole lung tissue)
    Gel Running Conditions
    Reduced Denaturing (4-15% gradient)
    Loading amount
    100 µg
    Specification
    RIPA lysate of whole lung tissue
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

    Ben Thomson

    Verified customer

    Submitted Aug 14 2015

    Application
    Western blot
    Loading amount
    40 µg
    Gel Running Conditions
    Reduced Denaturing (4-20% gradient gel)
    Sample
    Human Cell lysate - whole cell (HeLa cells)
    Specification
    HeLa cells
    Blocking step
    BSA as blocking agent for 2 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C

    Abcam user community

    Verified customer

    Submitted Jul 21 2014

    Abreviews
    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Blocking step
    BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 25°C
    Sample
    African Green Monkey Cell (COS7)
    Specification
    COS7
    Permeabilization
    Yes - Triton X 100 0.3%
    Fixative
    Paraformaldehyde

    Abcam user community

    Verified customer

    Submitted Jun 03 2013

    Answer

    Thank you for contacting us.

    I am sorry that this antibody did not perform as stated on the datasheet. Since this order was paid for by credit card, the refund will go back onto the card. To avoid confusion, please ensure your accounts department is aware of how the credit note is being used.

    Our accounting department can be contacted by email at us.credits@abcam.com or by telephone using the information at the Contact Us link in the top right corner of our website. Please refer to the credit note ID in any correspondence with our accounting department.

    The credit note ID is for your reference only and does not automatically guarantee the credit.

    I hope this experience will not prevent you from purchasing other products from us in the future. Our Scientific Support team is always at your service, should you require further expert advice.

    Read More

    Answer

    Thank you for taking time to contact us. I am sorry to hear this antibody is not providing satisfactory results.

    The details provided will enable us to investigate this case and will provide us with vital information for monitoring product quality. I appreciate the time you have spent in the laboratory and understand your concerns. It is regrettable the results have not been successful. Could I just ask to confirm the species that your samples were from (i.e. mouse)?

    As long as the antibody was being used in a tested species, I would be happy to offer you a replacment or refund. Please let me know which you would prefer.

    Thank you for your cooperation. I look forward to hearing from you with details of how you would like to proceed.

    Read More

    Answer

    I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab8227.

    To check the status of the order please contact our Customer Service team and reference this number.

    Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

    I wish you the best of luck with your research.

    Read More

    Answer

    Thank you for taking time to contact us. I am sorry to hear that this antibody is not providing satisfactory results.

    It sounds like you're seeing "inverse banding" which is white bands on a black background. Usually this means that you're using too much primary and/or secondary antibody. I'd recommend diluting the primary to 1:2000 and incubating this overnight at 4C in the blocking buffer (5% BSA) and using the secondary at 1:5000 - 10,000 for 1 hr at RT in blocking buffer. A 1:1000 dilution for a secondary antibody is quite high.

    Are you blocking in 5% BSA?

    How much protein are you loading and what are your samples (species, cell line, etc.)?

    Do you have an image that you can send?


    I would strongly recommend that you try diluting the primary and secondary more. Should the suggestions not improve the results, please do let me know.

    In the event that a product is not functioning in the species and applications cited on the product datasheet (and the problem has been reported within 6 months of purchase), we would be pleased to provide a free of charge replacement, credit note, or refund.

    I hope this information is helpful, and I thank you for your cooperation.

    Read More

    Answer

    Thank you very much for your reply.

    Please see below for the testing code to try ab121733 with zebrafish samples:

    DISCOUNT CODE: ***
    EXPIRATION DATE: December 12, 2012

    I do hope that this antibody will work well with zebrafish, but if you need any assitance optimizing the protocol or anything else, please let me know and I'll be happy to help.

    Have a nice day!

    Read More

    Answer

    Thank you very much for your reply.

    The testing codes can actuallybe used to receive any primary antibody in our catalog, regardless of target. So, if the WDR81 antibody does not work with zebrafish samples, you can use the code to get any other primary antibody of your choosing.

    I hope that this information will be useful, but please let me know if you have further questions. Have a nice day!

    Read More

    Answer

    Thank you very much for your email.

    The immunogen is only 46% identical to the zebrafish sequence, but there are a couple of areas of high homology, so it's hard to say whether this antibody might react with zebrafish or not. My guess is that the antibody will weakly cross-react, but you may need to use a higher concentration to see a signal. We unfortunately don't have any other WDR81 antibodies to compare it to.

    If you would like a discount code to try this antibody, please let me know and I will send one to you.

    I hope that this helps! Have a great weekend and I look forward to hearing from you soon.

    Read More

    1-10 of 40 Abreviews or Q&A

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