Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-GTPase HRAS antibody [Y132] (ab32417)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [Y132] to GTPase HRAS
  • Suitable for: WB, IP
  • Knockout validated
  • Reacts with: Mouse, Rat, Human

Overview

  • Product name

    Anti-GTPase HRAS antibody [Y132]
    See all GTPase HRAS primary antibodies

  • Description

    Rabbit monoclonal [Y132] to GTPase HRAS

  • Host species

    Rabbit

  • Specificity

    Reactivity with other RAS members has not been tested.

  • Tested Applications & Species

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

  • Immunogen

    Synthetic peptide within Human GTPase HRAS aa 150 to the C-terminus (C terminal). The exact sequence is proprietary.

  • Positive control

    • MCF7 and PC12 cell lysates and MCF7 cells.

  • General notes

     

     

    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-GTPase HRAS antibody [Y132] (ab32417)
    Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)
    All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/500 dilution

    Lane 1 : Wild-type HEK-293 whole cell lysate
    Lane 2 : HRAS knockout HEK-293 whole cell lysate
    Lane 3 : HeLa whole cell lysate
    Lane 4 : MCF7 whole cell lysate

    Lysates/proteins at 20 µg/ml per lane.


    Lanes 1 - 4: Merged signal (red and green). Green - ab32417 observed at 21 kDa. Red - loading control, ab8245, observed at 37 kDa.

    ab32417 was shown to recognize HRAS in wild-type HEK-293 cells as signal was lost at the expected MW in HRAS knockout cells. Additional cross-reactive bands were observed in the wild-type and knockout cells. Wild-type and HRAS knockout samples were subjected to SDS-PAGE. The membrane was blocked with 3% Milk. Ab32417 and ab8245 (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-GTPase HRAS antibody [Y132] (ab32417)
    Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)
    All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/2500 dilution (purified)

    Lane 1 : mouse brain lysate
    Lane 2 : rat brain lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Observed band size: 21 kDa
    why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)
    Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)
    All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/1000 dilution (purified)

    Lane 1 : MCF7 cell lysate
    Lane 2 : HeLa cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : HRP goat anti-rabbit IgG (H+L) at 1/20000 dilution

    Observed band size: 21 kDa why is the actual band size different from the predicted?



    Blocking buffer: 5% NFDM/TBST
    Dilution buffer: 5% NFDM/TBST

  • Immunoprecipitation - Anti-GTPase HRAS antibody [Y132] (ab32417)
    Immunoprecipitation - Anti-GTPase HRAS antibody [Y132] (ab32417)

    ab32417 (purified) at 1/60 immunoprecipitating GTPase in 10 μg mouse brain whole cell lysate (Lanes 1 and 2, observed at 21 kDa). Lane 3 - PBS. For western blotting, HRP Veriblot for IP Detection Reagent (ab131366) was used for detection (1/10 000). Blocking buffer and concentration: 5% NFDM/TBST Dilution buffer and concentration: 5% NFDM/TBST

  • Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)
    Western blot - Anti-GTPase HRAS antibody [Y132] (ab32417)
    All lanes : Anti-GTPase HRAS antibody [Y132] (ab32417) at 1/500 dilution (unpurified)

    Lane 1 : MCF7 cell lysate
    Lane 2 : PC12 cell lysate

    Observed band size: 21 kDa why is the actual band size different from the predicted?

  • Anti-GTPase HRAS antibody [Y132] (ab32417)
    Anti-GTPase HRAS antibody [Y132] (ab32417)

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