Recombinant
RabMAb

Recombinant Anti-Galectin 3 antibody [EP2775Y] - BSA and Azide free (ab197544)

Rabbit recombinant monoclonal Galectin 3 antibody [EP2775Y]. Validated in WB, IHC, Flow Cyt, ICC/IF and tested in Mouse, Rat, Human.

Overview

  • Product name

    Anti-Galectin 3 antibody [EP2775Y] - BSA and Azide free
    See all Galectin 3 primary antibodies
  • Description

    Rabbit monoclonal [EP2775Y] to Galectin 3 - BSA and Azide free
  • Host species

    Rabbit
  • Tested applications

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

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide within Human Galectin 3 aa 150-250. The exact sequence is proprietary.

  • Positive control

    • A375, HeLa, SW480 and A431 lysates. This antibody gave a positive result when used in the following formaldehyde fixed cell lines: Panc-1
  • General notes

    Ab197544 is the carrier-free version of ab76245. 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.

    ab197544 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.

Properties

Applications

Our Abpromise guarantee covers the use of ab197544 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
Flow Cyt Use at an assay dependent concentration.

ab199376 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

 

IHC-P Use at an assay dependent concentration.
WB Use at an assay dependent concentration. Predicted molecular weight: 26 kDa.
ICC/IF Use at an assay dependent concentration.
  • Application notes
    Is unsuitable for 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

    Images

    • Immunohistochemical analysis of formaldehyde fixed mouse lung tissue sections labelling Galectin 3 with ab76245 at a dilution of 1/6000. The secondary antibody used was biotin conjugated goat anti rabbit IgG at a dilution of 1/300. Antigen retrieval was heat mediated using citric acid.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76245).

    • Flow Cytometry analysis of HeLa (human cervix adenocarcinoma) cells labeling Galectin 3 with purified ab76245 at 1/50 dilution (10ug/mL) (red). Cells were fixed with 4% paraformaldehyde and permeabilised with 90% methanol. A Goat anti rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody. Rabbit monoclonal IgG (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 (ab76245).

    • Immunocytochemistry/Immunofluorescence analysis of HT-29 cells labelling Galectin with purified ab76245 at 1/1000. 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 (1/500) and ab150120, an Alexa Fluor® 594-conjugated goat anti-mouse IgG (1/500) 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 (ab76245).

    • ICC/IF image of unpurified ab76245 stained Panc-1 cells. The cells were 4% formaldehyde fixed (10 min) and then incubated in 1%BSA / 10% normal goat serum / 0.3M glycine in 0.1% PBS-Tween for 1h to permeabilise the cells and block non-specific protein-protein interactions. The cells were then incubated with the antibody ab76245 at 10µg/ml overnight at +4°C. The secondary antibody (pseudo-colored green) was Alexa Fluor® 488 goat anti- rabbit (ab150081) IgG (H+L) preadsorbed, used at a 1/1000 dilution for 1h. Alexa Fluor® 594 WGA was used to label plasma membranes (pseudo-colored red) at a 1/200 dilution for 1h at room temperature. DAPI was used to stain the cell nuclei (pseudo-colored blue) at a concentration of 1.43µM for 1hour at room temperature.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76245).

    • Overlay histogram showing THP1 cells stained with unpurified ab76245 (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 (ab76245, 1/1000 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) (0.1μg/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.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76245).

    • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human lung squamous carcinoma tissue labelling Galectin 3 with unpurified ab76245.

      This data was developed using the same antibody clone in a different buffer formulation containing PBS, BSA, glycerol, and sodium azide (ab76245).

    • This IHC data was generated using the same anti-Galectin 3 antibody clone [EP2775Y] in a different buffer formulation (cat# ab76245).

       Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of human thyroid carcinoma tissue labelling Galectin 3 with purified ab76245 at 1/250. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. ab97051, a HRP-conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody (1/500). Negative control using PBS instead of primary antibody. Counterstained with hematoxylin.

    References

    ab197544 has not yet been referenced specifically in any publications.

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