Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)

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 - BSA and Azide free
  • Suitable for: WB
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free
    See all alpha Actinin/ACTN1 primary antibodies

  • Description

    Rabbit monoclonal [EP2528Y] to alpha Actinin/ACTN1 - BSA and Azide free

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    WB
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.

  • Positive control

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

  • General notes

    ab185204 is the carrier-free version of ab81265 This format is designed for use in antibody labeling, including fluorochromes, metal isotopes, oligonucleotides, enzymes.

     

    Our carrier-free formats are supplied in a buffer free of BSA, sodium azide and glycerol for higher conjugation efficiency.

    Use our conjugation kits  for antibody conjugates that are ready-to-use in as little as 20 minutes with

    Ab185204 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

    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.

Properties

Images

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    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?



    This data was developed using the same antibody clone in a different buffer formulation (ab81265).

    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] - BSA and Azide free (ab185204)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)

    This data was developed using ab81265, the same antibody clone in a different buffer formulation.

    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] - BSA and Azide free (ab185204)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    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



    This data was developed using ab81265, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    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



    This data was developed using ab81265, the same antibody clone in a different buffer formulation.

    Blocking and dilution buffer: 5% NFDM/TBST.

  • Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    Western blot - Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    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



    This data was developed using ab81265, the same antibody clone in a different buffer formulation.

  • Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)
    Anti-alpha Actinin/ACTN1 antibody [EP2528Y] - BSA and Azide free (ab185204)

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