Validated using a knockout cell lineRecombinantRabMAb

Recombinant Anti-Galectin 3 antibody [EPR19244] (ab209344)

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Key features and details

  • Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
  • Rabbit monoclonal [EPR19244] to Galectin 3
  • Suitable for: IP, ICC/IF, Flow Cyt, IHC-P, WB
  • Knockout validated
  • Reacts with: Human

Overview

  • Product name

    Anti-Galectin 3 antibody [EPR19244]
    See all Galectin 3 primary antibodies

  • Description

    Rabbit monoclonal [EPR19244] to Galectin 3

  • Host species

    Rabbit

  • Tested applications

    Suitable for: IP, ICC/IF, Flow Cyt, IHC-P, WBmore details

  • Species reactivity

    Reacts with: Human

  • Immunogen

    Recombinant full length protein within Human Galectin 3 aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P17931

  • Positive control

    • WB: HeLa, SW480, MCF7 and A431 whole cell lysates; human heart, kidney and stomach lysates. IHC-P: Human liver, stomach, diffuse large B cell lymphoma and colon cancer tissues. ICC/IF: HeLa and A431 cells. Flow Cyt: HeLa cells. IP: HeLa whole cell lysate.

  • 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.

    Reproducibility is key to advancing scientific discovery and accelerating scientists’ next breakthrough.

    Abcam is leading the way with our range of recombinant antibodies, knockout-validated antibodies and knockout cell lines, all of which support improved reproducibility.

    We are also planning to innovate the way in which we present recommended applications and species on our product datasheets, so that only applications & species that have been tested in our own labs, our suppliers or by selected trusted collaborators are covered by our Abpromise™ guarantee.

    In preparation for this, we have started to update the applications & species that this product is Abpromise guaranteed for.

    We are also updating the applications & species that this product has been “predicted to work with,” however this information is not covered by our Abpromise guarantee.

    Applications & species from publications and Abreviews that have not been tested in our own labs or in those of our suppliers are not covered by the Abpromise guarantee.

    Please check that this product meets your needs before purchasing. If you have any questions, special requirements or concerns, please send us an inquiry and/or contact our Support team ahead of purchase. Recommended alternatives for this product can be found below, as well as customer reviews and Q&As.

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: 7.2
    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA
  • Concentration information loading...

  • Purity

    Protein A purified

  • Clonality

    Monoclonal

  • Clone number

    EPR19244

  • Isotype

    IgG

  • Research areas

Applications

Our Abpromise guarantee covers the use of ab209344 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
IP 1/30.
ICC/IF 1/500.
Flow Cyt 1/50.
IHC-P 1/2000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 26 kDa (predicted molecular weight: 26 kDa).

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

    Images

    • Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      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 - ab209344 observed at 35 kDa. Red - loading control, ab8245, observed at 37 kDa.

      ab209344 was shown to specifically react with Galectin 3 (KO) in wild-type HAP1 cells. No band was observed when LGALS3 (KO) knockout samples were examined. Wild-type and Galectin 3 (KO) knockout samples were subjected to SDS-PAGE. Ab209344 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 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.

    • Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Lanes 1-2 : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/10000 dilution
      Lanes 3-4 : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/1000 dilution

      Lane 1 : HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate
      Lane 2 : SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate
      Lane 3 : MCF7 (Human breast adenocarcinoma cell line) whole cell lysate
      Lane 4 : A431 (Human epidermoid carcinoma cell line) whole cell lysate

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

      Predicted band size: 26 kDa
      Observed band size: 26 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1/2: 30 seconds; Lane 3/4: 8 seconds

    • Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Western blot - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      All lanes : Anti-Galectin 3 antibody [EPR19244] (ab209344) at 1/1000 dilution

      Lane 1 : Human heart tissue
      Lane 2 : Human kidney tissue
      Lane 3 : Human stomach tissue

      Lysates/proteins at 10 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution

      Predicted band size: 26 kDa
      Observed band size: 26 kDa



      Blocking/Dilution buffer: 5% NFDM/TBST.

      Exposure time: Lane 1/2: 15 seconds; Lane 3: 30 seconds

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Positive staining on Kupffer cells in the liver is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on human stomach is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on human diffuse large B cell lymphoma is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Galectin 3 with ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on human colon cancer is observed. Counter stained with Hematoxylin.

      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/500 dilution.

      Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

    • Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 with ab209344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L(Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). 

      Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

      The negative controls are as follows:
      -ve control 1: ab209344 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
      -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L(Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunocytochemistry/ Immunofluorescence - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling Galectin 3 with ab209344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weakly cytoplasmic staining on A431 cell line. The nuclear counter stain is DAPI (blue). 

      Tubulin is detected with Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution (red).

      The negative controls are as follows:
      -ve control 1: ab209344 at 1/500 dilution followed by ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (ab150120) secondary antibody at 1/1000 dilution.
      -ve control 2: Anti-alpha Tubulin mouse MAb (ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • Flow Cytometry - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Flow Cytometry - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 with ab209344 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.

    • Immunoprecipitation - Anti-Galectin 3 antibody [EPR19244] (ab209344)
      Immunoprecipitation - Anti-Galectin 3 antibody [EPR19244] (ab209344)

      Galectin 3 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with ab209344 at 1/30 dilution. Western blot was performed from the immunoprecipitate using ab209344 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/10000 dilution.

      Lane 1: HeLa whole cell lysate 10µg (Input).

      Lane 2: ab209344 IP in HeLa whole cell lysate.

      Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab209344 in HeLa whole cell lysate.

      Blocking and dilution buffer and concentration: 5% NFDM/TBST.

      Exposure time: 8 seconds.

    References (0)

    Publishing research using ab209344? Please let us know so that we can cite the reference in this datasheet.

    ab209344 has not yet been referenced specifically in any publications.

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