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 (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.

Detection of Human Galectin-3 by Simple Western^TM.

Simple Western lane view shows lysates of Exosome Standards (Human Urine) (NBP2-49840) and MCF-7 human breast cancer cell line, loaded at 0.5 mg/ml. A specific band was detected for Galectin-3 at approximately 37 kDa (as indicated) using 10 µg/ml of Goat Anti-Human Galectin-3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1154) followed by HRP-conjugated Donkey Anti-Goat Secondary Antibody (Catalog # 042-206). This experiment was conducted under reducing conditions and using the 12-230kDa separation system.

Detection of Human Galectin‑3 by Simple Western^TM.

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.

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; (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 Western Blot

GPNMB and galectin-3 levels are elevated in FTD-GRN brains. a, b GPNMB and galectin-3 levels (ng/mg protein) were measured in frontal lobe tissue lysates generated from cognitively normal controls (CTL; n = 27) and FTD-GRN patients (n = 25). Data analyzed using unpaired t-test. c Representative immunoblots for GPNMB and galectin-3 in frontal lobe lysates from cognitively normal controls (n = 8) and FTD-GRN (n = 8) patients. d GPNMB levels (ng/mL) in CSF samples form cognitively normal controls (n = 14), FTD-GRN (n = 9), FTD-C9orf72 (n = 12) and FTD-MAPT (n = 12) samples quantified by ELISA. Data analyzed using one-way ANOVA. e, f GPNMB immunostaining was performed on frontal lobe tissue sections from cognitively normal controls (n = 5) (e) and FTD-GRN (n = 5) (f) patients. g, h Immunostaining for p-TDP 43 was stained on adjacent sections from identical samples in e, f as marker of FTLD pathology. i GPNMB staining intensity in human brain sections (e, f) were measured and presented as fold change. Representative immunofluorescence staining for cell markers (green) (j, n, r), GPNMB (red) (k, o, s), DAPI (blue) (i, p, t) in paraffin sections of brains from FTD-GRN cases. Iba-1, GFAP, NeuN used for markers of human microglia, astrocytes, and neurons respectively. GPNMB and Iba-1 signals overlap (arrow) (m) whereas, no overlapping signal was observed in co-staining with GFAP or NeuN (q, u). Scale bars were labeled in the images. Data analyzed by unpaired t-test. Scale bars (20 µm) labeled in images and quantitative data are shown as mean ± SEM, *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/33028409), licensed under a CC-BY license. Not internally tested by R&D Systems.

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Applications for Human Galectin-3 Antibody

Application

Recommended Usage

CyTOF-ready

Ready to be labeled using established conjugation methods. No BSA or other carrier proteins that could interfere with conjugation.

Immunohistochemistry

5-15 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human colon and lung cancer tissue

Intracellular Staining by Flow Cytometry

2.5 µg/10⁶ cells
Sample: Human PBMCs fixed with paraformaldehyde and permeabilized with saponin

Simple Western

10 µg/mL
Sample: Exosome Standards (Human Urine) (Catalog # NBP2-49840), COLO 205 human colorectal adenocarcinoma cell line, MCF‑7 human breast cancer cell line, and U‑118‑MG human glioblastoma/astrocytoma cell line

Western Blot

0.1 µg/mL
Sample: COLO 205 human colorectal adenocarcinoma cell line, MCF‑7 human breast cancer cell line, and U‑118‑MG human glioblastoma/astrocytoma cell line

Reviewed Applications

Read 3 reviews rated 4.7 using AF1154 in the following applications:

Imaging Mass Cytometry (1 Review)
Immunocytochemistry/Immunofluorescence (1 Review)
Immunofluorescence (1 Review)

Formulation, Preparation, and Storage

Purification

Antigen Affinity-purified

Reconstitution

Reconstitute at 0.2 mg/mL in sterile PBS. For liquid material, refer to CoA for concentration.

Formulation

Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. See Certificate of Analysis for details.
*Small pack size (-SP) is supplied either lyophilized or as a 0.2 µm filtered solution in PBS.

Shipping

Lyophilized product is shipped at ambient temperature. Liquid small pack size (-SP) is shipped with polar packs. Upon receipt, store immediately at the temperature recommended below.

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.

Background: Galectin-3

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 epsilonBP, 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).

References

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.

Alternate Names

AGE-R3, CBP35, GAL3, Galectin3, L29, LGALS3, Mac-2

Entrez Gene IDs

3958 (Human); 16854 (Mouse)

Gene Symbol

LGALS3

UniProt

P17931.5

Additional Galectin-3 Products

Product Documents for Human Galectin-3 Antibody

Product Datasheets

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COA

SDS

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Product Specific Notices for Human Galectin-3 Antibody

For research use only

Citations
Reviews

Calculators
Flow Cytometry

Human Galectin-3 Antibody

R&D Systems, part of Bio-Techne | Catalog # AF1154

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Scientific Data Images for Human Galectin-3 Antibody

Detection of Human Galectin‑3 by Western Blot.

Detection of Human Galectin-3 by Simple WesternTM.

Detection of Human Galectin‑3 by Simple WesternTM.