Validated using a knockout cell line
Recombinant
RabMAb

Anti-Galectin 3 antibody [EPR2774] (ab76466)

Overview

  • Product name
    Anti-Galectin 3 antibody [EPR2774]
    See all Galectin 3 primary antibodies
  • Description
    Rabbit monoclonal [EPR2774] to Galectin 3
  • Tested applications
    Suitable for: WB, Flow Cyt, ICC/IFmore details
    Unsuitable for: IHC-P or IP
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    A synthetic peptide corresponding to residues in human Galectin 3

  • Positive control
    • WB: HeLa, SW480, A431 and MCF7 cell lysates ICC/IF: HeLa cells.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

    Mouse: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

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

Properties

Applications

Our Abpromise guarantee covers the use of ab76466 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 1/2000 - 1/10000. Predicted molecular weight: 26 kDa.
Flow Cyt 1/30 - 1/100. ab172730-Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.
ICC/IF 1/100 - 1/250.
  • Application notes
    Is unsuitable for IHC-P or IP.
  • Target

    • Function
      Galactose-specific lectin which binds IgE. May mediate with the alpha-3, beta-1 integrin the stimulation by CSPG4 of endothelial cells migration. Together with DMBT1, required for terminal differentiation of columnar epithelial cells during early embryogenesis.
    • Tissue specificity
      A major expression is found in the colonic epithelium. It is also abundant in the activated macrophages.
    • Sequence similarities
      Contains 1 galectin domain.
    • Cellular localization
      Nucleus. Cytoplasmic in adenomas and carcinomas. May be secreted by a non-classical secretory pathway and associate with the cell surface.
    • Information by UniProt
    • Database links
    • Alternative names
      • 35 kDa lectin antibody
      • Carbohydrate binding protein 35 antibody
      • Carbohydrate-binding protein 35 antibody
      • CBP 35 antibody
      • CBP35 antibody
      • Gal-3 antibody
      • GAL3 antibody
      • Galactose-specific lectin 3 antibody
      • Galactoside-binding protein antibody
      • GALBP antibody
      • Galectin 3 internal gene,included antibody
      • Galectin-3 antibody
      • Galectin3 antibody
      • GALIG antibody
      • GBP antibody
      • IgE binding protein antibody
      • IgE-binding protein antibody
      • L 31 antibody
      • L 34 antibody
      • L-31 antibody
      • L-34 galactoside-binding lectin antibody
      • L31 antibody
      • Laminin-binding protein antibody
      • Lectin L-29 antibody
      • Lectin, galactose binding, soluble 3 antibody
      • LEG3_HUMAN antibody
      • LGALS2 antibody
      • LGALS3 antibody
      • MAC 2 antigen antibody
      • Mac-2 antibody
      • Mac-2 antigen antibody
      • MAC2 antibody
      • Macrophage galactose-specific lectin antibody
      • MGC105387 antibody
      see all

    Anti-Galectin 3 antibody [EPR2774] images



    • Predicted band size : 26 kDa
      Observed band size : 35 kDa (why is the actual band size different from the predicted?)

      Lane 1: Wild type HAP1 whole cell lysate (20 µg)
      Lane 2: Galectin 3 (KO) knockout HAP1 whole cell lysate (20 µg)
      Lane 3: HeLa whole cell lysate (20 µg)
      Lane 4: MCF7 whole cell lysate (20 µg)
      Lanes 1 - 4: Merged signal (red and green). Green - ab76466 observed at 35 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab76466 was shown to specifically react with Galectin 3 (KO) when Galectin 3 (KO) knockout samples were used. Wild-type and Galectin 3 (KO) knockout samples were subjected to SDS-PAGE. Ab76466 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 2000 dilution and 1/10000 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.


    • Overlay histogram showing THP1 cells stained with ab76466 (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% human serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab76466, 1/100 dilution) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) (ab150077) at 1/2000 dilution for 30 min at 22°C. Isotype control antibody (black line) was rabbit IgG (monoclonal) (1μg/1x106 cells) used under the same conditions. Unlabelled sample (blue line) was also used as a control. Acquisition of >5,000 events were collected using a 20mW Argon ion laser (488nm) and 525/30 bandpass filter.
    • ab76466, at a 1/100 dilution, staining Galectin 3 in HeLa cells by Immunofluorescence.
    • All lanes : Anti-Galectin 3 antibody [EPR2774] (ab76466) at 1/5000 dilution

      Lane 1 : HeLa cell lysate
      Lane 2 : SW480 cell lysate
      Lane 3 : A431 cell lysate
      Lane 4 : MCF7 cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      goat anti-rabbit HRP at 1/2000 dilution

      Predicted band size : 26 kDa
      Observed band size : 30 kDa (why is the actual band size different from the predicted?)

    References for Anti-Galectin 3 antibody [EPR2774] (ab76466)

    ab76466 has not yet been referenced specifically in any publications.

    Product Wall

    Abcam has not validated the combination of species/application used in this Abreview.
    Application
    Western blot
    Sample
    Mouse Cell lysate - whole cell (BMDM)
    Gel Running Conditions
    Reduced Denaturing (4-12% Bis Tris gel, MES running buffer)
    Loading amount
    5 µg
    Specification
    BMDM
    Blocking step
    Licor TBS blocking buffer as blocking agent for 15 minute(s) · Concentration: 100% · Temperature: 22°C
    Username

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    Verified customer

    Submitted Aug 24 2016

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

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