The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Use a concentration of 1 - 5 µg/ml.
Use a concentration of 5 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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.
Peripheral blood leukocytes and lymphatic tissues. Overexpressed in Hodgkin disease tissue.
Contains 2 galectin domains.
Contains two homologous but distinct carbohydrate-binding domains.
Cytoplasm. Secreted. May also be secreted by a non-classical secretory pathway.
IHC image of galectin 9 staining in Human normal Tonsil formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab153673, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-galectin 9 antibody [1G3] (ab153673)
All lanes : Anti-galectin 9 antibody [1G3] (ab153673) at 5 µg/ml
Lane 1 : HeLa transfected with Galectin 9 whole cell lysate Lane 2 : HeLa (wild type control) whole cell lysate
Lysates/proteins at 10 µg per lane.
Secondary All lanes : Goat anti-mouse IgG polyclonal antibody (HRP conjugated) at 1/10000 dilution
Developed using the ECL technique.
Performed under reducing conditions.
Additional bands at: 18 kDa, 35 kDa. We are unsure as to the identity of these extra bands.
Barjon C et al. A novel monoclonal antibody for detection of galectin-9 in tissue sections: application to human tissues infected by oncogenic viruses. Infect Agent Cancer7:16 (2012).
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