Overview

  • Product name
    Anti-alpha Actinin antibody [AT6/172]
    See all alpha Actinin primary antibodies
  • Description
    Mouse monoclonal [AT6/172] to alpha Actinin
  • Host species
    Mouse
  • Specificity
    Recognizes and is specific for human alpha Actinin 1 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
    Suitable for: WB, IHC-Fr, IHC (Methanol fixed), IHC-P, ICC/IFmore details
    Unsuitable for: IP
  • Species reactivity
    Reacts with: Mouse, Sheep, Rabbit, Human
  • Immunogen

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

  • Positive control
    • WB: Human pancreatic cancer whole cell lysate. ICC/IF: HeLa cells.

Properties

Applications

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.

Application Abreviews Notes
WB Use a concentration of 0.5 - 2 µg/ml. Predicted molecular weight: 103 kDa.
IHC-Fr Use a concentration of 15 µg/ml.
IHC (Methanol fixed) Use a concentration of 15 µg/ml.
IHC-P Use at an assay dependent concentration.
ICC/IF Use a concentration of 15 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

    • Anti-alpha Actinin antibody [AT6/172] (ab18061) at 1/2000 dilution + Human pancreatic cancer cell line - whole cell lysate at 20 µg

      Secondary
      HRP conjugated goat anti-mouse antibody at 1/2000 dilution

      Developed using the ECL technique.

      Performed under reducing conditions.

      Predicted band size: 103 kDa
      Observed band size: 103 kDa


      Exposure time: 1 minute

      See Abreview

    • 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.

    • ab18061 staining alpha Actinin in Mouse skin tumor tissue sections by Immunohistochemistry (IHC-P - paraformaldehyde-fixed, paraffin-embedded sections). Tissue was fixed with paraformaldehyde and blocked with 5% serum for 1 hour at 20°C; antigen retrieval was by heat mediation in a citrate buffer. Samples were incubated with primary antibody (1/50) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated Donkey anti-mouse polyclonal (1/400) was used as the secondary antibody.

      See Abreview

    References

    This product has been referenced in:
    • Zheng C  et al. Gastrodin Inhibits Store-Operated Ca(2+) Entry and Alleviates Cardiac Hypertrophy. Front Pharmacol 8:222 (2017). ICC/IF ; Mouse . Read more (PubMed: 28487655) »
    • Han X  et al. Zyxin regulates endothelial von Willebrand factor secretion by reorganizing actin filaments around exocytic granules. Nat Commun 8:14639 (2017). Read more (PubMed: 28256511) »
    See all 15 Publications for this product

    Customer reviews and Q&As

    1-10 of 11 Abreviews or Q&A

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (mouse skin tumor)
    Specification
    mouse skin tumor
    Fixative
    Paraformaldehyde
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: 10mm sodium citrate PH 6.0
    Permeabilization
    No
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C

    Abcam user community

    Verified customer

    Submitted Feb 15 2013

    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
    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

    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

    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
    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Sheep Tissue lysate - whole (Whole Placentome)
    Loading amount
    100 µg
    Specification
    Whole Placentome
    Gel Running Conditions
    Reduced Denaturing (10% SDS PAGE)
    Blocking step
    Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

    Dr. Bryan White

    Verified customer

    Submitted May 11 2011

    Answer

    Thank you for your enquiry. The immunogen for ab18061 is full-length human alpha-actinin 1 from platelets. There is no epitope information for this monoclonal antibody; however, the sequence homology between human, mouse and rat alpha-actinin 1 is 99% for all combinations. I hope this information helps you, and please do not hesitate to contact us if you need any more advice or information.

    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 (Lung carcinoma)
    Loading amount
    40 µg
    Specification
    Lung carcinoma
    Gel Running Conditions
    Reduced Denaturing (12%)
    Blocking step
    Milk as blocking agent for 16 hour(s) and 0 minute(s) · Concentration: 2% · Temperature: 4°C

    Abcam user community

    Verified customer

    Submitted Apr 24 2009

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Western blot
    Sample
    Human Cell lysate - whole cell (pancreatic cancer cell line)
    Loading amount
    20 µg
    Specification
    pancreatic cancer cell line
    Gel Running Conditions
    Reduced Denaturing (4-12%)
    Blocking step
    Milk as blocking agent for 30 minute(s) · Concentration: 5% · Temperature: 22°C

    Abcam user community

    Verified customer

    Submitted Jul 09 2008

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunocytochemistry/ Immunofluorescence
    Sample
    Human Cell (cardiomyocyte)
    Specification
    cardiomyocyte
    Fixative
    Paraformaldehyde
    Blocking step
    Serum as blocking agent for 18 hour(s) and 0 minute(s) · Concentration: 10%

    Abcam user community

    Verified customer

    Submitted Feb 09 2007

    1-10 of 11 Abreviews or Q&A

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

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