Recombinant
RabMAb

Recombinant Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046)

Overview

  • Product name

    Anti-galectin 9/Gal-9 antibody [EPR22214]
    See all galectin 9/Gal-9 primary antibodies
  • Description

    Rabbit monoclonal [EPR22214] to galectin 9/Gal-9
  • Host species

    Rabbit
  • Specificity

    This antibody is not recommended for mouse and rat in IHC.

  • Tested applications

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

    Reacts with: Human
  • Immunogen

    Synthetic peptide within Human galectin 9/Gal-9 aa 250-350. The exact sequence is proprietary.
    Database link: O00182

  • Positive control

    • IHC-P: Human liver and colon tissue. WB: U937, HL-60 and THP-1 whole cell lysates. ICC/IF: U937 cells. Flow Cyt: U937 cells.
  • 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

Applications

Our Abpromise guarantee covers the use of ab227046 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 1/1000. Predicted molecular weight: 40 kDa.
IHC-P 1/1000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
ICC/IF 1/250.
Flow Cyt 1/500.
  • Application notes
    Is unsuitable for IP.
  • Target

    • Function

      Binds galactosides. Has high affinity for the Forssman pentasaccharide. May play a role in thymocyte-epithelial interactions relevant to the biology of the thymus. Inhibits cell proliferation. It is a ligand for HAVCR2/TIM3. Induces T-helper type 1 lymphocyte (Th1) death. Isoform Short acts as an eosinophil chemoattractant.
    • Tissue specificity

      Peripheral blood leukocytes and lymphatic tissues. Overexpressed in Hodgkin disease tissue.
    • Sequence similarities

      Contains 2 galectin domains.
    • Domain

      Contains two homologous but distinct carbohydrate-binding domains.
    • Cellular localization

      Cytoplasm. Secreted. May also be secreted by a non-classical secretory pathway.
    • Information by UniProt
    • Database links

    • Alternative names

      • 36 kDa beta-galactoside-binding lectin antibody
      • Ecalectin antibody
      • Gal-9 antibody
      • galectin 9 antibody
      • Galectin-9 antibody
      • galectin9 antibody
      • HOM HD 21 antibody
      • HOMHD21 antibody
      • HUAT antibody
      • Lectin galactoside binding soluble 9 antibody
      • LEG9_HUMAN antibody
      • LGAL S9 antibody
      • LGALS 9 antibody
      • Lgals9 antibody
      • LGALS9A antibody
      • MGC117375 antibody
      • MGC125973 antibody
      • MGC125974 antibody
      • Tumor antigen HOM-HD-21 antibody
      • UAT antibody
      • Urate transporter/channel antibody
      • Urate transporter/channel protein antibody
      see all

    Images

    • Immunohistochemical analysis of paraffin-embedded human liver tissue labeling galectin 9/Gal-9 with ab227046 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear and cytoplasmic staining on Kupffer cells of human liver (PMID: 20209097, PMID: 18202194) is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized U937 (human histiocytic lymphoma cell line) cells labeling galectin 9/Gal-9 with ab227046 at 1/250 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 in U-937 cells. The nuclear counterstain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at 1/200 dilution (red). 

      Secondary antibody only control: Used PBS instead of primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

      PANC-1 (human pancreatic epithelial cancinoma cell line) cells stained in the same manner serve as a negative control.

    • All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution

      Lane 1 : U937 (human histiocytic lymphoma monocyte), whole cell lysate
      Lane 2 : HL-60 (human Acute Promyelocytic Leukemia promyeloblast), whole cell lysate
      Lane 3 : MOLT-4 (human lymphoblastic leukemia T lymphoblast), whole cell lysate
      Lane 4 : PANC-1 (human pancreatic epithelioid carcinoma epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 40 kDa
      Observed band size: 34-39 kDa
      why is the actual band size different from the predicted?
      Additional bands at: 13-15 kDa (possible degradation product)


      Exposure time: 59 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).

      Degraded fragments (13-15 kDa) have also been descried (PMID: 15811318).

      Negative control: PANC-1.

    • Flow cytometric analysis of 4% paraformaldehyde-fixed, 90% methanol permeabilized PANC-1 (human pancreatic epithelial cancinoma cell line) (Left) / U937 (human histiocytic lymphoma cell line) (Right) cell line labeling galectin 9/Gal-9 with ab227046 at 1/500 (red) compared with a Rabbit monoclonal IgG (ab172730) (black) and an unlabeled control (cells without incubation with primary antibody and secondary antibody) (blue). Goat anti rabbit IgG (Alexa Fluor® 488, ab150077), at 1/2000 dilution was used as the secondary antibody.

      Negative Control - PANC-1 cells.

    • Immunohistochemical analysis of paraffin-embedded human colon tissue labeling galectin 9/Gal-9 with ab227046 at 1/1000 dilution, followed by a ready to use Goat Anti-Rabbit IgG H&L (HRP). Nuclear and cytoplasmic staining on epithelial cells of human colon (PMID: 18202194) is observed. Counterstained with hematoxylin.
      Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is a ready to use Goat Anti-Rabbit IgG H&L (HRP).
      Perform heat mediated antigen retrieval using ab93684 (Tris/EDTA buffer, pH 9.0).

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HEK-293T (human epithelial cell line from embryonic kidney) cells labeling galectin 9/Gal-9 with ab227046 at 1/250 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

      Confocal image showing no staining in HEK-293T cells

      Negative control: HEK-293T (PMID: 11698107). The nuclear counterstain is DAPI (blue).
      Counterstained with ab195889 Anti-alpha Tubulin antibody [DM1A] - Microtubule Marker (Alexa Fluor® 594) at a 1/200 dilution (red).
      Secondary antibody only control: Used PBS instead of the primary antibody, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution.

    • All lanes : Anti-galectin 9/Gal-9 antibody [EPR22214] (ab227046) at 1/1000 dilution

      Lane 1 : THP-1 (human monocytic leukemia monocyte), whole cell lysate
      Lane 2 : HEK-293T (human embryonic kidney epithelial cell), whole cell lysate

      Lysates/proteins at 20 µg per lane.

      Secondary
      All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution

      Predicted band size: 40 kDa
      Observed band size: 34-39 kDa why is the actual band size different from the predicted?
      Additional bands at: 13-15 kDa (possible degradation product)


      Exposure time: 59 seconds


      Blocking/Dilution buffer: 5% NFDM/TBST.

      The expression of the isoforms observed is consistent with what has been described in the literature (PMID 11886844, PMID: 22805533).

      Degraded fragments (13-15 kDa) has also been described (PMID: 15811318).

      Negative control: HEK-293T (PMID: 24333696).

    References

    ab227046 has not yet been referenced specifically in any publications.

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