Validated using a knockout cell line
Recombinant
RabMAb

Anti-N WASP antibody [EPR6959] (ab126626)

Overview

  • Product name
    Anti-N WASP antibody [EPR6959]
    See all N WASP primary antibodies
  • Description
    Rabbit monoclonal [EPR6959] to N WASP
  • Tested applications
    Suitable for: Flow Cyt, WB, IHC-P, ICC/IFmore details
  • Species reactivity
    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide, corresponding to a region within Human N WASP.

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

    This product is a recombinant rabbit monoclonal antibody.

     

    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

Applications

Our Abpromise guarantee covers the use of ab126626 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 at an assay dependent concentration.

Purified format

WB 1/1000 - 1/10000. Detects a band of approximately 65 kDa (predicted molecular weight: 55 kDa).
IHC-P 1/50 - 1/100. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF 1/300.

For unpurified, use 1/50 - 1/100.

Target

  • Function
    Regulates actin polymerization by stimulating the actin-nucleating activity of the Arp2/3 complex. Binds to HSF1/HSTF1 and forms a complex on heat shock promoter elements (HSE) that negatively regulates HSP90 expression.
  • Sequence similarities
    Contains 1 CRIB domain.
    Contains 1 WH1 domain.
    Contains 2 WH2 domains.
  • Cellular localization
    Cytoplasm > cytoskeleton. Nucleus. Preferentially localized in the cytoplasm when phosphorylated and in the nucleus when unphosphorylated.
  • Information by UniProt
  • Database links
  • Alternative names
    • DKFZp779G0847 antibody
    • MGC48327 antibody
    • N-WASP antibody
    • Neural Wiskott Aldrich syndrome protein antibody
    • Neural Wiskott-Aldrich syndrome protein antibody
    • NWASP antibody
    • Wasl antibody
    • WASL_HUMAN antibody
    • WASPB antibody
    • Wiskott Aldrich syndrome gene like antibody
    • Wiskott Aldrich syndrome gene like protein antibody
    • Wiskott Aldrich syndrome like antibody
    • WiskottAldrich syndrome like antibody
    see all

Images



  • Predicted band size : 55 kDa

    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.

  • 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 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.

  • 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.
  • 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?)
  • 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
    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

  • 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
    Goat anti-rabbit HRP conjugated antibody at 1/2000 dilution

    Predicted band size : 55 kDa
  • Unpurified ab126626, at 1/50, staining N WASP in formalin fixed, paraffin embedded human papillary carcinoma tissue by Immunohistochemistry

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

References

This product has been referenced in:
  • Bendris N  et al. SNX9 promotes metastasis by enhancing cancer cell invasion via differential regulation of RhoGTPases. Mol Biol Cell N/A:N/A (2016). Read more (PubMed: 26960793) »
  • Saengsawang W  et al. CIP4 coordinates with phospholipids and actin-associated proteins to localize to the protruding edge and produce actin ribs and veils. J Cell Sci 126:2411-23 (2013). Read more (PubMed: 23572514) »

See all 3 Publications for this product

Customer reviews and Q&As

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Mouse Tissue sections (Pancreas and Pancreatic tumor (PDAC))
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: High pH
Permeabilization
No
Specification
Pancreas and Pancreatic tumor (PDAC)
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 18°C
Fixative
Paraformaldehyde
Username

Clara Lubeseder-Martellato

Verified customer

Submitted Jan 24 2017

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Mouse Tissue lysate - whole (Pancreas)
Gel Running Conditions
Reduced Denaturing (10%)
Loading amount
30 µg
Specification
Pancreas
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
Username

Clara Lubeseder-Martellato

Verified customer

Submitted Jun 09 2016

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

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