Overview

  • Product name

    Anti-alpha Actinin/ACTN1 antibody [MAC 276]
    See all alpha Actinin/ACTN1 primary antibodies
  • Description

    Rat monoclonal [MAC 276] to alpha Actinin/ACTN1
  • Host species

    Rat
  • Tested applications

    Suitable for: Flow Cyt, ICC/IF, IHC-P, Electron Microscopy, WBmore details
  • Species reactivity

    Reacts with: Rat, Chicken, Human, Drosophila melanogaster, Waterbug
  • Immunogen

    Tissue, cells or virus corresponding to Waterbug alpha Actinin/ACTN1.

  • Positive control

    • In Western Blot, this antibody gave a positive signal in human and rat skeletal muscle tissue lysates, and in rat heart tissue lysate. This antibody gave a positive result in IHC in the following FFPE tissue: Human normal bladder.
  • General notes

     This product was previously labelled as alpha Actinin

     

Properties

Applications

Our Abpromise guarantee covers the use of ab50599 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 1µg for 106 cells.

ab18450 - Rat monoclonal IgG2a, is suitable for use as an isotype control with this antibody.

 

ICC/IF Use a concentration of 5 µg/ml.
IHC-P Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval via the pressure cooker method before commencing with IHC staining protocol.
Electron Microscopy Use at an assay dependent concentration.
WB Use a concentration of 1 µg/ml. Detects a band of approximately 103 kDa (predicted molecular weight: 103 kDa).

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

  • All lanes : Anti-alpha Actinin/ACTN1 antibody [MAC 276] (ab50599) at 1 µg/ml

    Lane 1 : Human skeletal muscle tissue lysate - total protein (ab29330)
    Lane 2 : Heart (Rat) Tissue Lysate
    Lane 3 : Skeletal Muscle (Rat) Tissue Lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Peroxidase Conjugated AffiniPure Rabbit Anti-Rat IgG (H+L) at 1/10000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

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


    Exposure time: 8 minutes
  • IHC image of alpha Actin staining in Human normal bladder formalin fixed paraffin embedded tissue section. The section was pre-treated using pressure cooker heat mediated antigen retrieval with sodium citrate buffer (pH6) for 20 mins at 98°C. The section was incubated with ab50599, 5µg/ml, for 15 mins at room temperature. A Goat anti-Rat biotinylated secondary antibody was used to detect the primary, and visualized using an HRP conjugated ABC system. The section was counterstained with haematoxylin and mounted with DPX.

  • ICC/IF image of ab68826 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 ab50599 5µg/ml overnight at +4°C. The secondary antibody (green) was Alexa Fluor® 488 goat anti- rat 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.

  • Overlay histogram showing HeLa cells stained with ab50599 (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 (ab50599, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rat IgG (H+L) (ab98386) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rat IgG2a, kappa monoclonal [aRTK2758] (ab18450, 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/permeabilized in 0.1% PBS-Tween used under the same conditions.

References

This product has been referenced in:

  • Valarmathi MT  et al. Functional Tissue Engineering: A Prevascularized Cardiac Muscle Construct for Validating Human Mesenchymal Stem Cells Engraftment Potential In Vitro. Tissue Eng Part A 24:157-185 (2018). Read more (PubMed: 28457188) »
  • Xiang Q  et al. ISL1 overexpression enhances the survival of transplanted human mesenchymal stem cells in a murine myocardial infarction model. Stem Cell Res Ther 9:51 (2018). Read more (PubMed: 29482621) »
See all 7 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Mouse Cell (cardiomyocytes)
Permeabilization
Yes - Triton X-100
Specification
cardiomyocytes
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Jun 23 2014

Question
Answer

The guarantee is a promise of a replacement, credit, or refund if an antibody fails in an application listed on the datasheet, with samples derived from the species listed in the "Cross-reactivity" field of the datasheet. For ab2787, as an example, the guarantee applies to:

Application: ICC, IHC-P, IP, WB

Species: Human, Mouse, Rat, Cow, Fruit fly (Drosophila melanogaster), Rabbit, Saccharomyces cerevisiae and Xenopus laevis

The species listed as "predicted" are not covered by the guarantee. The guarantee is in effect for 6 months following purchase.

The following link has a fuller description:

www.abcam.com/abpromise

If you have any questions, please contact me.

Read More

Answer

Thank you for sending the fruit fly sequences. Alignment of the following two sequences with the immunogens fo the antibodies we have against the human targets did not reveal any significant homology, (i.e., none greater than 50% identity) and I am skeptical that they will be able to detect the fly proteins.

Human name= SQSTM1 / p62 Fly name= Ref(2)P (CG10360)

Human name = BAG3/CAIR1/BIS Fly name=Starvin/BAG3 (CG32130)

The HSP70 antibodies that list Drosophila and IHC in the respective fields on the datasheet (Cross reactivity; Application) will be covered by our guarantee, even though we do no have data on the datasheet. The absence of data may be a consequence of the fact that we source many of these antibodies from other laboratories that state the results but do not provide the data or references.

To choose the likeliest to work, I suggest an antibody that has the most references, for instance ab2787 or ab5442.

Read More

Question
Answer

Please find below the discount code we discussed for ab50599. Please send the Drosophila SQSTM1 / p62 sequence at your convenience and I will align it with the ab56416 immunogen sequence.

I am very pleased to hear you would like to accept our offer and test ab50599 in IHC of Drosophila flight muscle. This code will give you 1 free primary antibody before the expiration date. To redeem this offer, please submit an Abreview for IHC (you may need to use the IHC review form for frozen tissue (IHC-Fr)). Please include this code in the “Additional Comments” section so we know the Abreview is for this promotion. For more information on how to submit an Abreview, please visit the site: www.abcam.com/Abreviews.


We publish both positive and negative Abreviews on our datasheets so please submit the results of your tests regardless of the outcome. The code will be active once the Abreview has been submitted and can be redeemed in one of the following ways: 1) Call to place your order and mention the code to our customer service department; 2) Include the code in your fax order; 3) Place your order on the web and enter the promotional code.

Any feedback that you can provide will be greatly appreciated, whether positive or negative. If you have any further questions, please do not hesitate to contact us. We look forward to receiving your Abreview and wish you luck with your research.

The terms and conditions applicable to this offer can be found here: www.abcam.com/collaborationdiscount.

Read More
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Cardiomyocytes and fibroblasts (from ES cell diff))
Permeabilization
Yes - 0.1% Triton-X-100
Specification
Cardiomyocytes and fibroblasts (from ES cell diff)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Fixative
Methanol

Abcam user community

Verified customer

Submitted Apr 20 2010

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
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