Overview

  • Product name

    Anti-Moesin antibody [EP1863Y] - BSA and Azide free
    See all Moesin primary antibodies
  • Description

    Rabbit monoclonal [EP1863Y] to Moesin - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-FoFr, WB, IP, IHC-P, Flow Cyt, ICC/IFmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Moesin aa 450-550 (C terminal). The exact sequence is proprietary.
    Database link: P26038
    (Peptide available as ab201545)

  • Positive control

    • WB: Wild-type and Moesin knockout HAP1, HeLa and Raji whole cell lysate.
  • General notes

    Ab232580 is the carrier-free version of ab52490. 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.

    ab232580 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

  • Form

    Liquid
  • Storage instructions

    Shipped at 4°C. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle.
  • Storage buffer

    pH: 8
    Constituent: PBS
  • Concentration information loading...
  • Purity

    Protein A purified
  • Clonality

    Monoclonal
  • Clone number

    EP1863Y
  • Isotype

    IgG
  • Research areas

Applications

Our Abpromise guarantee covers the use of ab232580 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
IHC-FoFr Use at an assay dependent concentration. PubMed: 19853564
WB Use at an assay dependent concentration. Predicted molecular weight: 68 kDa.
IP Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
Flow Cyt Use at an assay dependent concentration.
ICC/IF Use at an assay dependent concentration.

Target

  • Function

    Probably involved in connections of major cytoskeletal structures to the plasma membrane.
  • Tissue specificity

    In all tissues and cultured cells studied.
  • Sequence similarities

    Contains 1 FERM domain.
  • Post-translational
    modifications

    Phosphorylation on Thr-558 is crucial for the formation of microvilli-like structures.
  • Cellular localization

    Cell membrane. Cytoplasm > cytoskeleton. Apical cell membrane. Cell projection > microvillus membrane. Phosphorylated form is enriched in microvilli-like structures at apical membrane (By similarity). Increased cell membrane localization of both phosphorylated and non-phosphorylated forms seen after thrombin treatment.
  • Information by UniProt
  • Database links

  • Alternative names

    • Epididymis luminal protein 70 antibody
    • HEL70 antibody
    • Membrane organizing extension spike protein antibody
    • Membrane-organizing extension spike protein antibody
    • MOES_HUMAN antibody
    • Moesin antibody
    • Moesin/anaplastic lymphoma kinase fusion protein antibody
    • Msn antibody
    • MSN/ALK fusion antibody
    see all

Images

  • Lane 1: Wild-type HAP1 whole cell lysate (20 µg)
    Lane 2: Moesin knockout HAP1 whole cell lysate (20 µg)
    Lane 3: HeLa whole cell lysate (20 µg)
    Lane 4: Raji whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab52490 observed at 75 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab52490 was shown to specifically react with Moesin in wild-type HAP1 cells as signal was lost in Moesin knockout cells. Wild-type and Moesin knockout samples were subjected to SDS-PAGE. ab52490 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at 1/1,000 dilution and 1/20,000 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 1/20,000 dilution for 1 hour 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 (ab52490).

     

  • ab52490 (purified) at 1/20 immunoprecipitating Moesin in HeLa whole cell lysate. 10 ug of cell lysate was present in the input. For western blotting, a HRP-conjugated Veriblot for IP Detection Reagent (ab131366) (1/10,000) was used for detection. A rabbit monoclonal IgG (ab172730) was used intead of ab128913 as a negative control (Lane 3).

    Blocking buffer and concentration: 5% NFDM/TBST.

    Diluting buffer and concentration: 5% NFDM /TBST.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52490).

  • Flow Cytometry analysis of HeLa cells labelling Moesin with purified ab52490 at 1/30 (red). Cells were fixed with 4% paraformaldehyde. A FITC-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. Black - Isotype control, rabbit monoclonal IgG. Blue - Unlabelled control, cells without incubation with primary and secondary antibodies.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52490).

  • Immunocytochemistry/Immunofluorescence analysis of HeLa (human cervix adenocarcinoma) cells labelling Moesin with purified ab52490 at 1/100. Cells were fixed with 4% paraformaldehyde and permeabilized with 0.1% Triton X-100. ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500) was used as the secondary antibody. DAPI (blue) was used as the nuclear counterstain. ab7291, a mouse anti-tubulin (1/1000) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/1000) were also used.

    Control 1: primary antibody (1/100) and secondary antibody, ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500).

    Control 2: ab7291 (1/1000) and secondary antibody, ab150077, an Alexa Fluor® 488-conjugated goat anti-rabbit IgG (1/500).

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52490).

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human spleen tissue labelling Moesin with purified ab52490 at 1/50. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a goat anti-rabbit IgG H&L (HRP) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52490).

  • Overlay histogram showing HeLa cells stained with unpurified ab52490 (red line). The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab52490, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-rabbit IgG (H+L) (ab96899) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed.

    This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab52490).

References

ab232580 has not yet been referenced specifically in any publications.

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