Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)

Reviews (9)Q&A (6)References (25)
Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)
  • Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)

Key features and details

  • Mouse monoclonal [AT6/172] to alpha Actinin/ACTN1
  • Suitable for: ICC
  • Reacts with: Human
  • Isotype: IgG1

Overview

  • Product name

    Anti-alpha Actinin/ACTN1 antibody [AT6/172]
    See all alpha Actinin/ACTN1 primary antibodies

  • Description

    Mouse monoclonal [AT6/172] to alpha Actinin/ACTN1

  • Host species

    Mouse

  • Specificity

    Recognizes and is specific for human alpha Actinin 1/ACTN1 isoform (non-muscle) at 100 kDa.

    This antibody may recognize other isoforms of alpha-actinin (1-4). There is no specific epitope information available.

    The clone number has been updated from (0.T.02) to (AT6/172) both clone numbers name the same antibody clone.

  • Tested Applications & Species

    Application Species
    ICC
    Human
    See all applications and species data

  • Immunogen

    Full length protein corresponding to Human alpha Actinin/ACTN1.
    Database link: P12814

  • Positive control

    • ICC: HeLa cells, ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612).

  • General notes

     This product was previously labelled as alpha Actinin

     

Properties

Applications

The Abpromise guarantee

Our Abpromise guarantee covers the use of ab18061 in the following tested applications.

The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.

Guaranteed

Tested applications are guaranteed to work and covered by our Abpromise guarantee.

Predicted

Predicted to work for this combination of applications and species but not guaranteed.

Incompatible

Does not work for this combination of applications and species.

Application Species
ICC
Human
Application Abreviews Notes
ICC
Use a concentration of 1 - 5 µg/ml.
Notes
ICC
Use a concentration of 1 - 5 µg/ml.
  • Application notes
    Is unsuitable for IP.

    • Target

    • Function

      F-actin cross-linking protein which is thought to anchor actin to a variety of intracellular structures. This is a bundling protein.

    • Sequence similarities

      Belongs to the alpha-actinin family.
      Contains 1 actin-binding domain.
      Contains 2 CH (calponin-homology) domains.
      Contains 2 EF-hand domains.
      Contains 4 spectrin repeats.

    • Cellular localization

      Cytoplasm > cytoskeleton. Cytoplasm > myofibril > sarcomere > Z line. Colocalizes with MYOZ2 and PPP3CA at the Z-line of heart and skeletal muscle.
    • Information by UniProt

    • Database links

      • Alternative names

        • actinin 1 smooth muscle antibody
        • Actinin alpha 1 antibody
        • actinin, alpha 1 antibody
        • ACTN 1 antibody
        • Actn1 antibody
        • ACTN1_HUMAN antibody
        • Alpha Actinin 1 antibody
        • Alpha actinin cytoskeletal isoform antibody
        • Alpha-actinin cytoskeletal isoform antibody
        • Alpha-actinin-1 antibody
        • BDPLT15 antibody
        • F actin cross linking protein antibody
        • F-actin cross-linking protein antibody
        • FLJ40884 antibody
        • FLJ54432 antibody
        • Non muscle alpha actinin 1 antibody
        • Non-muscle alpha-actinin-1 antibody
        see all

      Images

      • Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)
        Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)

        Immunofluorescence staining of Actinin/ACTN1 using ab18061 in ioSkeletal Myocytes - Human iPSC-Derived Skeletal Myocytes (ab277612), which were differentiated for 10 days post induction.


        The cells were fixed with 100% MeOH (5 min) and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated overnight at +4°C with ab18061 at 5 µg/mL and ab6046, rabbit polyclonal to beta Tubulin, at 1/1000 dilution. Cells were then incubated with ab150117, Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed at 1/1000 dilution (shown in green) and ab150088, Goat Anti-Rabbit IgG H&L (Alexa Fluor® 594) preadsorbed at 1/1000 dilution (shown in red). Nuclear DNA was labelled with DAPI (shown in blue).


        Image was acquired with a high-content analyser (Operetta CLS, Perkin Elmer) and a maximum intensity projection of confocal sections is shown. Gamma is adjusted to 1.5 in all channels.

      • Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)
        Immunocytochemistry - Anti-alpha Actinin/ACTN1 antibody [AT6/172] (ab18061)

        ICC/IF image of ab18061 stained HeLa (Human epithelial cell line from cervix adenocarcinoma) cells. The cells were fixed in 4% formaldehyde for 10 minutes and then incubated in 1% BSA / 10% normal goat serum / 0.3 M glycine in 0.1% PBS-Tween for 1 hour to permeabilize the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody (ab18061, 1 µg/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 1 hour. Alexa Fluor® 594 WGA was used to label plasma membranes (red) at a 1/200 dilution for 1 hour. DAPI was used to stain the cell nuclei (blue) at a concentration of 1.43 µM.

      References (25)

      Publishing research using ab18061? Please let us know so that we can cite the reference in this datasheet.

      ab18061 has been referenced in 25 publications.

      • Tousley A  et al. Huntingtin associates with the actin cytoskeleton and a-actinin isoforms to influence stimulus dependent morphology changes. PLoS One 14:e0212337 (2019). PubMed: 30768638
      • Miki T  et al. Induced Pluripotent Stem Cell Derivation and Ex Vivo Gene Correction Using a Mucopolysaccharidosis Type 1 Disease Mouse Model. Stem Cells Int 2019:6978303 (2019). PubMed: 31065277
      • Tousley A  et al. Rac1 Activity Is Modulated by Huntingtin and Dysregulated in Models of Huntington's Disease. J Huntingtons Dis 8:53-69 (2019). PubMed: 30594931
      • Oury J  et al. MACF1 links Rapsyn to microtubule- and actin-binding proteins to maintain neuromuscular synapses. J Cell Biol 218:1686-1705 (2019). PubMed: 30842214
      • van den Dries K  et al. Modular actin nano-architecture enables podosome protrusion and mechanosensing. Nat Commun 10:5171 (2019). PubMed: 31729386
      View all Publications for this product

      Customer reviews and Q&As

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      1-10 of 15 Abreviews or Q&A

      Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha Actinin/ACTN1 antibody [AT6/172]

       Good
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Mouse Cell (Mouse vascular smooth muscle cells)
      Permeabilization
      Yes - NP40
      Specification
      Mouse vascular smooth muscle cells
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
      Fixative
      Formaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Oct 06 2020

      Immunocytochemistry/ Immunofluorescence abreview for Anti-alpha Actinin/ACTN1 antibody [AT6/172]

       Good
      Abreviews
      abreview image
      Application
      Immunocytochemistry/ Immunofluorescence
      Sample
      Human Cell (U2OS)
      Permeabilization
      Yes - NP40
      Specification
      U2OS
      Blocking step
      BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 3% · Temperature: 21°C
      Fixative
      Formaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Oct 06 2020

      Western blot abreview for Anti-alpha Actinin/ACTN1 antibody [AT6/172]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Mouse Cell lysate - whole cell (MC3T3 cells)
      Gel Running Conditions
      Reduced Denaturing
      Loading amount
      30 µg
      Specification
      MC3T3 cells
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      Submitted Sep 02 2020

      Western blot abreview for Anti-alpha Actinin/ACTN1 antibody [AT6/172]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Human Cell lysate - whole cell (U2OS)
      Gel Running Conditions
      Reduced Denaturing
      Loading amount
      25 µg
      Specification
      U2OS
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
      Read More

      Abcam user community

      Verified customer

      Submitted Aug 14 2020

      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) abreview for Anti-alpha Actinin/ACTN1 antibody [AT6/172]

       Good
      Abreviews
      abreview image
      Application
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
      Sample
      Mouse Tissue sections (mouse skin tumor)
      Antigen retrieval step
      Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6.0
      Permeabilization
      No
      Specification
      mouse skin tumor
      Blocking step
      Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
      Fixative
      Paraformaldehyde
      Read More

      Abcam user community

      Verified customer

      Submitted Feb 15 2013

      Question

      Thank you for your email - the problem is that we used same conditions as before (the replaced antibody with long incubation time), however, we did not get any SUR1 bands at all. You think 1:100 dilution is OK to use, but we incubated the blot at this concentration for very long time (3 days), this is quite different from the most other antibodies people used! This time even incubating for 3 days won't work (but beta actin is fine).

      We do not know what else we could do?

      Read More
      Answer

      We now have new lot in stock so I can send you one vial as a replacement antibody. Again because we haven't tested this lot in mouse so we cannot confirm the ab will work.

      You have kindly mentioned in previous emails that the antibody worked before so perhaps running suggested human lysates will give goodindication whether the antibody is wrong or the lysates.

      Let me know if you are interested in getting the new lot.

      Read More

      Question

      Please see attached - let me know if you need any further info.
      thanks,

      Read More
      Answer

      Thank you for confirming all the details.

      The mouse species has been added to the datasheet from a publication http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?cmd=Retrieve&db=PubMed&list_uids=20362548&dopt=Abstract. We unfortunately hold no tested data with mouse lysates. The mouse and human SUR1 are quite similar at the immunogenic region so it is more likely that the antibody will work.

      We currently haven't received any complaint other than the 2 you submitted last year. So we are not sure if this issue is lot related. I have checked the case again and would like to suggest changing few things and hoping that the antibody will be OK;

      - SUR1 is multipass membrane protein; for mmp we do not recommend boiling the sample with loading buffer. Please try heating samples for 60-70C for 10-15 minutes.
      - The lot conc. we have sent is 0.5 mg/ml. 1/100 dilution is OK to use.
      - if possible please include a positive control with human cell lines e.g. U2OS, A431, pancreatic tissue lysates, Tonsil, gall bladder, liver or lung tissue lysates.

      Please address all the emails of this complaint to me (ab32844).

      Let me know how these suggestions go. if the results does not improve I will then ask to choose another anti SUR1 antibody.

      Read More

      Question

      Thank you for your email.

      Previously we had difficulty with the ab32844 anti-SUR1 antibody (last year), and the replaced antibody was working OK for both WB and ICC (using longer time for incubation in WB, similar to this time, we could detect the expected band). It used up quickly, but that gave us some useful WB data.

      We currently need to do more WB in some sets of samples for a manuscript, and hope the newly ordered SUR1 antibody will work (but this lot seems not working).

      Please let me know if you need any other information.

      Thank you again,

      Read More
      Answer

      Thank you for contacting us.

      I am sorry to hear you are experiencing difficulties with one of our products. We take product complaints very seriously, and investigate every product that we feel may not be performing correctly.

      I am attaching our questionnaire so that we can gather further information regarding the samples tested and the protocol used. Once we receive the completed questionnaire, we will look at the protocol and see if there are any suggestions we can make that may improve the results. This information will also allow us to investigate this case internally and initiate additional testing where necessary. If the product was purchased in the last six months and is being used according to our Abpromise, we would be happy to replace or refund the antibody.

      I look forward to receiving your reply.

      PS:Please provide all the details of experiemtn andany troubleshooting you have already done! Thanks. Please do not forget to add the purchase order number.

      Read More

      Question

      I would like to ask you for help. We need a specific antibody only labeling cardiomyocytes. If possible, the antibody should be labeled already. I have already bought the TroponinI antibody (ab97711) of Abcam which is working quite well. If you could offer another one as desribed, I would be glad. Thank you in advance for your help! Kind regards

      Read More
      Answer

      Thank you for contacting us. There are many proteins that act as markers for Cardiomyocytes; antibodies against these targets can be used for ICC/IF or other required applications. Main cardiac proteins are as follows; - alpha actin; alpha sarcomeric Actin antibody [Alpha Sr-1] (ab28052) - N-Cadherin: N Cadherin antibody (ab12221) - alpha actinin; alpha Actinin antibody [0.T.02] (ab18061) - bFGF: FGF basic antibody (ab16828) I would recommend typing the target name in the search box provided on the Abcam homepage and then selecting the antibodies as per your requirements e.g. monoclonal, polyclonal, rabbit or mouse etc. The full list of Cardiomyocytes markers can be found by clicking the below link; http://www.antibodybeyond.com/reviews/cell-markers/cardiomyocyte-marker.htm Please click on the following link to explore the Abcam Cardiovascular page https://www.abcam.com/index.html?pageconfig=resource&rid=10694 I hope this information is helpful to you. Please do not hesitate to contact us if you need any more advice or information.

      Read More

      Western blot abreview for Anti-alpha Actinin/ACTN1 antibody [AT6/172]

       Excellent
      Abreviews
      abreview image
      Application
      Western blot
      Sample
      Sheep Tissue lysate - whole (Whole Placentome)
      Gel Running Conditions
      Reduced Denaturing (10% SDS PAGE)
      Loading amount
      100 µg
      Specification
      Whole Placentome
      Blocking step
      Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C
      Read More

      Dr. Bryan White

      Verified customer

      Submitted May 11 2011

      1-10 of 15 Abreviews or Q&A

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