Validated using a knockout cell line
Recombinant
RabMAb

Recombinant Anti-N WASP antibody [EPR6959] - BSA and Azide free (ab232457)

Overview

  • Product name

    Anti-N WASP antibody [EPR6959] - BSA and Azide free
    See all N WASP primary antibodies
  • Description

    Rabbit monoclonal [EPR6959] to N WASP - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: WB, IHC-P, ICC/IF, Flow Cytmore details
  • Species reactivity

    Reacts with: Mouse, Human
  • Immunogen

    Synthetic peptide within Human N WASP aa 300-450. The exact sequence is proprietary.
    Database link: O00401

  • Positive control

    • WB: A549, wild-type and N WASP knockout HAP1 cell lysate; Human thyroid tissue lysate.
  • General notes

    Ab232457 is the carrier-free version of ab126626. 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 <1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.

    ab232457 is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm.

    Maxpar® is a trademark of Fluidigm Canada Inc.

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

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab232457 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
WB Use at an assay dependent concentration. Detects a band of approximately 65 kDa (predicted molecular weight: 55 kDa).
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
ICC/IF Use at an assay dependent concentration.
Flow Cyt Use at an assay dependent concentration.

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

  • 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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126626).

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

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126626).

  • Equilibrium disassociation constant (KD)
    Learn more about KD

    Click here to learn more about KD

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (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.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab126626).

     

References

ab232457 has not yet been referenced specifically in any publications.

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