Astana Biomed Group, an authorized Abcam distributor in Central Asia

Anti-N WASP antibody [EPR6959] (ab126626)

Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR6959] to N WASP
  • Suitable for: Flow Cyt, WB, IHC-P, ICC/IF
  • Knockout validated
  • Reacts with: Mouse, Human

Overview

  • Product name

    Anti-N WASP antibody [EPR6959]
    See all N WASP primary antibodies

  • Description

    Rabbit monoclonal [EPR6959] to N WASP

  • Host species

    Rabbit

  • Tested Applications & Species

    Application Species
    Flow Cyt
    Human
    ICC/IF
    Human
    IHC-P
    Human
    WB
    Mouse
    Human
    See all applications and species data

  • Immunogen

    Synthetic peptide within Human N WASP aa 300-450. The exact sequence is proprietary.

  • Positive control

    • NIH 3T3, F9, L929, HeLa and K562 cell lysates; Human papillary carcinoma tissue.

  • 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-N WASP antibody [EPR6959] (ab126626)
    Western blot - Anti-N WASP antibody [EPR6959] (ab126626)

    Lane 1: Wild-type HAP1 cell lysate (20 µg)
    Lane 2: N WASP knockout HAP1 cell lysate (20 µg)
    Lane 3: A549 cell lysate (20 µg)
    Lane 4: Human thyroid tissue lysate (20 µg)
    Lanes 1 - 4: Merged signal (red and green). Green - ab126626 observed at 67 kDa. Red - loading control, ab8245, observed at 37 kDa.
    ab126626 was shown to recognize N WASP when N WASP knockout samples were used, along with additional cross-reactive bands. Wild-type and N WASP knockout samples were subjected to SDS-PAGE. ab126626 and ab8245 (loading control to GAPDH) were diluted 1/1000 and 1/2000 respectively and incubated overnight at 4°C. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed (ab216776) secondary antibodies at 1/10000 dilution for 1 h at room temperature before imaging.

  • Immunocytochemistry/ Immunofluorescence - Anti-N WASP antibody [EPR6959] (ab126626)
    Immunocytochemistry/ Immunofluorescence - Anti-N WASP antibody [EPR6959] (ab126626)

    Immunofluorescence staining of K562 cells with purified ab126626 at a working dilution of 1/300, counter-stained with DAPI. The secondary antibody was Alexa Fluor® 488 goat anti-rabbit (ab150077), used at a dilution of 1/1000. ab7291, a mouse anti-tubulin antibody (1/1000), was used to stain tubulin along with ab150120 (Alexa Fluor® 594 goat anti-mouse, 1/1000), shown in the top right hand panel. The cells were fixed in 4% PFA and permeabilized using 0.1% Triton X 100. The negative controls are shown in bottom middle and right hand panels - for negative control 1, purified ab126626 was used at a dilution of 1/500 followed by an Alexa Fluor® 594 goat anti-mouse antibody (ab150120) at a dilution of 1/500. For negative control 2, ab7291 (mouse anti-tubulin) was used at a dilution of 1/500 followed by an Alexa Fluor® 488 goat anti-rabbit antibody (ab150077) at a dilution of 1/400.

  • Flow Cytometry - Anti-N WASP antibody [EPR6959] (ab126626)
    Flow Cytometry - Anti-N WASP antibody [EPR6959] (ab126626)

    Flow Cytometry analysis of K562 (human chronic myelogenous leukemia) cells labeling N WASP with purified ab126626 at 1/150 (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) was used as the secondary antibody. Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730) (Black) was used as the isotype control, cells without incubation with primary antibody and secondary antibody (Blue) were used as the unlabeled control.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N WASP antibody [EPR6959] (ab126626)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N WASP antibody [EPR6959] (ab126626)
    Immunohistochemical staining of paraffin embedded human kidney with purified ab126626 at a working dilution of 1/100. The secondary antibody used is ab97051, a goat anti-rabbit IgG (H&L) at a dilution of 1/500. The sample is counter-stained with hematoxylin. Antigen retrieval was perfomed using Tris-EDTA buffer, pH 9.0. PBS was used instead of the primary antibody as the negative control, and is shown in the inset.
  • Western blot - Anti-N WASP antibody [EPR6959] (ab126626)
    Western blot - Anti-N WASP antibody [EPR6959] (ab126626)
    Anti-N WASP antibody [EPR6959] (ab126626) at 1/10000 dilution (purified) + L929 whole cell lysate at 10 µg

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

    Predicted band size: 55 kDa
    Observed band size: 65 kDa
    why is the actual band size different from the predicted?

  • Western blot - Anti-N WASP antibody [EPR6959] (ab126626)
    Western blot - Anti-N WASP antibody [EPR6959] (ab126626)
    All lanes : Anti-N WASP antibody [EPR6959] (ab126626) at 1/10000 dilution (purified)

    Lane 1 : HeLa whole cell lysate
    Lane 2 : K562 whole cell lysate

    Lysates/proteins at 10 µg per lane.

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

    Predicted band size: 55 kDa
    Observed band size: 65 kDa why is the actual band size different from the predicted?



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

  • Western blot - Anti-N WASP antibody [EPR6959] (ab126626)
    Western blot - Anti-N WASP antibody [EPR6959] (ab126626)
    All lanes : Anti-N WASP antibody [EPR6959] (ab126626) at 1/1000 dilution (Unpurified)

    Lane 1 : NIH 3T3 cell lysate
    Lane 2 : F9 cell lysate
    Lane 3 : L929 cell lysate
    Lane 4 : HeLa cell lysate
    Lane 5 : K562 cell lysate

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size: 55 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N WASP antibody [EPR6959] (ab126626)
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-N WASP antibody [EPR6959] (ab126626)

    Unpurified ab126626, at 1/50, staining N WASP in formalin fixed, paraffin embedded human papillary carcinoma tissue by Immunohistochemistry

    Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.

  • OI-RD Scanning - Anti-N WASP antibody [EPR6959] (ab126626)
    OI-RD Scanning - Anti-N WASP antibody [EPR6959] (ab126626)
    Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD
  • Anti-N WASP antibody [EPR6959] (ab126626)
    Anti-N WASP antibody [EPR6959] (ab126626)

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