Validated using a knockout cell line
Recombinant
RabMAb

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

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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

     

     

    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

    Preservative: 0.01% Sodium azide
    Constituents: 59% PBS, 40% Glycerol, 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

    ab209344 has not yet been referenced specifically in any publications.

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

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