Detects human Galectin-3 in direct ELISAs and Western blots. In direct ELISAs, approximately 50% cross-reactivity with recombinant mouse Galectin-3 is observed and less than 1% cross-reactivity with recombinant human (rh) Galectin-1, rhGalectin-2, rhGalectin-4, rhGalectin-7, and rhGalectin-8 is observed.
Polyclonal Goat IgG
E. coli-derived recombinant human Galectin‑3 Ala2-Ile250 Accession # P17931.5
Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.
Intracellular Staining by Flow Cytometry
2.5 µg/106 cells
Human PBMCs fixed with paraformaldehyde and permeabilized with saponin
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Detection of Human Galectin‑3 by Western Blot. Western blot shows lysates of COLO 205 human colorectal adenocarcinoma cell line, MCF‑7 human breast cancer cell line, and U‑118‑MG human glioblastoma/astrocytoma cell line. PVDF membrane was probed with 0.1 µg/mL of Goat Anti-Human Galectin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1154) followed by HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF017). A specific band was detected for Galectin‑3 at approximately 28 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Galectin‑3 in Human Colon. Galectin‑3 was detected in formalin fixed paraffin-embedded sections of human colon using Goat Anti-Human Galectin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1154) at 1.7 µg/mL overnight at 4 °C. Tissue was stained using the Anti-Goat HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS008) and counterstained with hematoxylin (blue). View our protocol for Chromogenic IHC Staining of immersion fixed paraffin-embedded Tissue Sections.
Detection of Human Galectin‑3 by Simple WesternTM. Simple Western lane view shows lysates of COLO 205 human colorectal adenocarcinoma cell line, MCF‑7 human breast cancer cell line, and U‑118‑MG human glioblastoma/astrocytoma cell line, loaded at 0.2 mg/mL. A specific band was detected for Galectin‑3 at approximately 37-38 kDa (as indicated) using 10 µg/mL of Goat Anti-Human Galectin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1154) followed by 1:50 dilution of HRP-conjugated Anti-Goat IgG Secondary Antibody (Catalog # HAF109). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system.
Preparation and Storage
Reconstitute at 0.2 mg/mL in sterile PBS.
The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
Stability & Storage
Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
12 months from date of receipt, -20 to -70 °C as supplied.
1 month, 2 to 8 °C under sterile conditions after reconstitution.
6 months, -20 to -70 °C under sterile conditions after reconstitution.
The galectins constitute a large family of carbohydrate-binding proteins with specificity for N-acetyl-lactosamine-containing glycoproteins. At least 14 mammalian galectins, which share structural similarities in their carbohydrate recognition domains (CRD), have been identified. The galectins have been classified into the prototype galectins (-1, -2, -5, -7, -10, -11, -13, -14), which contain one CRD and exist either as a monomer or a noncovalent homodimer; the chimera galectins (Galectin-3) containing one CRD linked to a nonlectin domain; and the tandem-repeat galectins (-4, -6, -8, -9, -12) consisting of two CRDs joined by a linker peptide. Galectins lack a classical signal peptide and can be localized to the cytosolic compartments where they have intracellular functions. However, via one or more as yet unidentified non-classical secretory pathways, galectins can also be secreted to function extracellularly. Individual members of the galectin family have different tissue distribution profiles and exhibit subtle differences in their carbohydrate-binding specificities. Each family member may preferentially bind to a unique subset of cell-surface glycoproteins (1-4). Galectin-3, also known as Mac-2, L29, CBP35, and epsilon BP, is a chimera galectin that has a tendency to dimerize. Besides the soluble protein, alternatively spliced forms of chicken Galectin-3 containing a transmembrane-spanning domain and a leucine zipper motif have been reported. Galectin-3 is expressed in tumor cells, macrophages, activated T cells, osteoclasts, epithelial cells, and fibroblasts. It binds various matrix glycoproteins including laminin, fibronectin, LAMPS, 90K/Mac-2BP, MP20, and CEA. Galectin-3 promotes cell growth and proliferation for many cell types. Galectin-3 acts intracellularly to prevent apoptosis. Depending on the cell types, Galectin-3 exhibits pro- or anti-adhesive properties. Galectin-3 has proinflammatory activities in vitro and in vivo. It induces pro-inflammatory and inhibits Th2 type cytokine production. Galectin-3 chemoattracts monocytes and macrophages. It activates and degranulates basophils and mast cells. Elevated circulating levels of Galectin-3 has been show to correlate with the malignant potential of several types of cancer, suggesting that Galectin-3 is also involved in tumor growth and metastasis. Human and mouse Galectin-3 shares approximately 80% amino acid sequence similarity (1-5).
Rabinovich, A. et al. (2002) Trends in Immunol. 23:313.
Rabinovich, A. et al. (2002) J. Leukocyte Biology 71:741.
Hughes, R.C. (2001) Biochimie 83:667.
R&D Systems Cytokine Bulletin; Summer 2002.
Gorski, J.P. et al. (2002) J. Biol. Chem. 277:18840.
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