Anti-alpha Actinin antibody [MAC 276] (ab50599)

Overview

  • Product name
    Anti-alpha Actinin antibody [MAC 276]
    See all alpha Actinin primary antibodies
  • Description
    Rat monoclonal [MAC 276] to alpha Actinin
  • 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

    Flight muscle extract from Lethocerus indicus (Waterbug)

  • 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.

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

  • 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.

  • All lanes : Anti-alpha Actinin 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
    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

References

This product has been referenced in:
  • Sehring IM  et al. An equatorial contractile mechanism drives cell elongation but not cell division. PLoS Biol 12:e1001781 (2014). ICC/IF ; Sea tunicate . Read more (PubMed: 24503569) »
  • Chechenova MB  et al. The Drosophila Z-disc protein Z(210) is an adult muscle isoform of Zasp52, which is required for normal myofibril organization in indirect flight muscles. J Biol Chem 288:3718-26 (2013). IHC-Fr ; Drosophila melanogaster . Read more (PubMed: 23271733) »

See all 4 Publications for this product

Customer reviews and Q&As

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

Abcam user community

Verified customer

Submitted Jun 23 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (Cardiomyocytes and fibroblasts (from ES cell diff))
Specification
Cardiomyocytes and fibroblasts (from ES cell diff)
Fixative
Methanol
Permeabilization
Yes - 0.1% Triton-X-100
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 4% · Temperature: 25°C
Username

Abcam user community

Verified customer

Submitted Apr 20 2010

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

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