Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-alpha Actinin/ACTN1 antibody [EP2528Y]
    See all alpha Actinin/ACTN1 primary antibodies

  • Description

    Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Mouse
    Rat
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human alpha Actinin/ACTN1 aa 750-850 (C terminal). The exact sequence is proprietary.

  • Positive control

    • WB: HAP1, A431, NIH 3T3, C2C12, C6 and A673 cell lysates and human skeletal muscle and human heart tissue lysates.

  • General notes

     

     

     This product was previously labelled as alpha Actinin

     

    This product is a recombinant monoclonal antibody, which offers several advantages including:

    • - High batch-to-batch consistency and reproducibility
    • - Improved sensitivity and specificity
    • - Long-term security of supply
    • - Animal-free production
    For more information see here.

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

Properties

Images

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution

    Lane 1 : Wild-type HAP1 cell lysate
    Lane 2 : ACTN1 knockout HAP1 cell lysate
    Lane 3 : A431 cell lysate
    Lane 4 : Daudi cell lysate

    Lysates/proteins at 20 µg per lane.

    Performed under reducing conditions.

    Predicted band size: 103 kDa
    Observed band size: 115 kDa
    why is the actual band size different from the predicted?



    Lanes 1 - 4: Merged signal (red and green). Green - ab81265 observed at 115 kDa. Red - loading control ab7291 (Mouse anti-Alpha Tubulin [DM1A]) observed at 55kDa.

    ab81265 was shown to react with ACTN1 in wild-type HAP1 cells in western blot with loss of signal observed in ACTN1 knockout sample. Wild-type and ACTN1 knockout HAP1 cell lysates were subjected to SDS-PAGE. Membranes were blocked in fluorescent western blot (TBS-based) blocking solution before incubation with ab81265 and ab7291 (Mouse anti-Alpha Tubulin [DM1A]) overnight at 4°C at a 1 in 1000 Dilution and a 1 in 20000 dilution respectively. Blots were incubated with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

    Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Alpha Actinin knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: NIH 3T3 whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab81265 observed at 103 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab81265 was shown to recognize alpha Actinin in wild-type HAP1 cells as signal was lost at the expected MW in alpha Actinin knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and alpha Actinin knockout samples were subjected to SDS-PAGE. ab81265 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/500 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (purified)

    Lane 1 : NIH/3T3 whole cell lysate
    Lane 2 : C2C12 whole cell lysate
    Lane 3 : C6 whole cell lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (purified) + A673 whole cell lysate at 20 µg

    Secondary
    Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

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



    Blocking and dilution buffer: 5% NFDM/TBST
  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    All lanes : Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265) at 1/1000 dilution (unpurified)

    Lane 1 : NIH 3T3 cell lysate
    Lane 2 : human skeletal muscle tissue lysate
    Lane 3 : human heart tissue lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP-conjugated goat anti-rabbit IgG at 1/2000 dilution

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

  • Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)
    Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

Please note: All products are "FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC PROCEDURES"
For licensing inquiries, please contact partnerships@abcam.com

Alternative products to Anti-alpha Actinin/ACTN1 antibody [EP2528Y] (ab81265)

© 2021 Astana Biomed Group LLP. All rights reserved.