Human Galectin-3 Antibody (194805)

R&D Systems | Catalog # MAB11541

R&D Systems

Key Product Details

Species Reactivity

Validated:

Human

Cited:

Human

Applications

Validated:

Immunohistochemistry, Western Blot, ELISA Capture (Matched Antibody Pair), Simple Western

Cited:

Western Blot

Label

Unconjugated

Antibody Source

Monoclonal Mouse IgG2B Clone # 194805
View all Galectin-3 Primary Antibodies »

Product Specifications

Immunogen

E. coli-derived recombinant human Galectin‑3
Ala2-Ile250
Accession # P17931.5

Specificity

Detects human Galectin-3 in ELISAs. In sandwich immunoassays, no cross-reactivity with recombinant human Galectin-1, 2, 4, 7, 8, or 10 is observed.

Clonality

Monoclonal

Host

Mouse

Isotype

IgG2B

Scientific Data Images for Human Galectin-3 Antibody (194805)

Detection of Human Galectin‑3 antibody by Western Blot.

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 0.2 µg/mL of Mouse Anti-Human Galectin-3 Monoclonal Antibody (Catalog # MAB11541) followed by HRP-conjugated Anti-Mouse IgG Secondary Antibody (Catalog # HAF018). 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 antibody in Human Prostate Cancer Tissue by Immunohistochemistry (IHC-P).

Galectin‑3 in Human Prostate Cancer Tissue.

Galectin-3 was detected in immersion fixed paraffin-embedded sections of human prostate cancer tissue using Mouse Anti-Human Galectin-3 Monoclonal Antibody (Catalog # MAB11541) at 1.7 µg/mL for 1 hour at room temperature followed by incubation with the Anti-Mouse IgG VisUCyte™ HRP Polymer Antibody (Catalog # VC001). Tissue was stained using DAB (brown) and counterstained with hematoxylin (blue). Specific staining was localized to cytoplasm and plasma membrane in epithelial cells. View our protocol for IHC Staining with VisUCyte HRP Polymer Detection Reagents.
Detection of Human Galectin-3 antibody by Simple WesternTM.

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 38 kDa (as indicated) using 10 µg/mL of Mouse Anti-Human Galectin‑3 Monoclonal Antibody (Catalog # MAB11541). This experiment was conducted under reducing conditions and using the 12-230 kDa separation system. Non-specific interaction with the 230 kDa Simple Western standard may be seen with this antibody.
Human Galectin-3 ELISA Standard Curve

Human Galectin-3 ELISA Standard Curve

Recombinant Human Galectin‑3 (Catalog # 1154-GA) was serially diluted and captured by Mouse Anti-Human Galectin‑3 Monoclonal Antibody (Catalog # MAB11541) coated on a Clear Polystyrene Microplate (Catalog # DY990). Goat Anti-Human Galectin‑3 Antigen Affinity-purified Polyclonal Antibody (Catalog # AF1154) was biotinylated and incubated with the protein captured on the plate. Detection of the standard curve was achieved by incubating Streptavidin-HRP (Catalog # DY998)
Detection of Galectin-3 by Western Blot

Detection of Galectin-3 by Western Blot

FOXD1 is a transcription factor of Gal-3. (A) CL1-0 cells were co-transfected with plasmids of the Gal-3 promoter reporter (pGL3-Gal-3 2000 bp) or Gal-3 promoter mutation reporter (pGL3-Gal-3 2000 bp-mut) and FOXD1 (pFOXD1). The cell lysates were collected, and the promoter activity of Gal-3 was analyzed by luciferase assay. (B) ChIP-qPCR assay using IgG as a control or FOXD1 antibody in CL1-0 and CL1-5 showed the binding of FOXD1 on the Gal-3 promoter. (C) CL1-0 cells transfected with the empty vector or plasmid of FOXD1 (pFOXD1) for 48 h and (D) CL1-5 cells transfected with scramble or siRNA targeting FOXD1 (si-FOXD1) for 72 h were collected. The mRNA and protein levels of FOXD1 and Gal-3 were analyzed by qPCR (left) and immunoblotting (right), respectively. (E) CCK-8 assay in different groups. FOXD1-stimulated CL1-0 proliferation ability was reduced by knocking down Gal-3. (F) The colony-formation ability of FOXD1-stimulated CL1-0 cells was suppressed by the depletion of Gal-3. (G) The numbers of migrating or invading cells were increased in the FOXD1 group compared with the FOXD1-stimulated CL1-0-siGal-3 group and control group. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31795213), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Galectin-3 by Western Blot

Detection of Galectin-3 by Western Blot

Gal-3 regulates FOXD1 expression through integrin-beta 1 (ITG beta 1) signaling. (A,B) CL1-0 and CL1-5 cells were transfected with the empty vector or plasmid of Gal-3 (pGal-3) for 48 h or scramble and siRNA targeting Gal-3 (si-Gal-3) for 72 h, respectively, as indicated. The mRNA and protein levels of FOXD1 and Gal-3 were analyzed by qPCR and immunoblotting. CL1-5 cells were transfected with scramble and siRNA targeting Gal-3 for 72 h or empty vector or plasmid of Gal-3 for 48 h, respectively, as indicated, and immunoprecipitation was analyzed by immunoblotting. Input, the whole cell lysates. (C) CL1-0 cells were transfected with scramble, Gal-3, ITG beta 1 or si-Gal-3, and si-ITG beta 1 for 72 h, as indicated. Cells lysates were collected and analyzed by immunoblotting. (D) CL1-0 cells were transfected with Gal-3 or ITG beta 1 expression vector for 48 h, and immunoprecipitation was analyzed by immunoblotting. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31795213), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Galectin-3 by Western Blot

Detection of Galectin-3 by Western Blot

High FOXD1 and Gal-3 expression are associated with the poor prognosis in lung cancer. (A) The ranking of candidate molecules and the corresponding z-score and p-value using the Ingenuity Pathway Analyses (IPA) database from a microarray experiment performed with CL1-0 FOXD1 compared to CL1-0 vector and CL1-5 scramble compared to CL1-5 siRNA targeting FOXD1 (si-FOXD1) using a 5-fold change cut-off. (B) Statistical p-values and fold change of the expression of Gal-3 (LGALS3) and its relative molecules are shown. Prediction legend: upregulation (red round icon), downregulation (green round icon), predicted activation (brown round icon), predicted inhibition (blue round icon), leads to activation (brown line), leads to inhibition (blue line), findings inconsistent with stage of downstream molecule (yellow line), effect not predicted (gray line). (C) RT-qPCR analysis of Gal-3 and FOXD1 mRNA expression in various lung cancer cell lines. GAPDH was used as an internal control for mRNA loading. (D) Western blot analysis of Gal-3 and FOXD1 protein expression in various lung cancer cell lines. GAPDH was used as an internal control for protein loading. (E) Lung cancer patients with high gene level transcription of Gal-3 or FOXD1 expression level show a correlation with high risk and (F) poor disease-free survival. The data were retrieved and analyzed from the TCGA-LUAD samples (n = 475) and lung meta-base (n = 1044) of the SurvExpress database (http://bioinformatica.mty.itesm.mx:8080/Biomatec/SurvivaX.jsp). Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31795213), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Galectin-3 by Western Blot

Detection of Galectin-3 by Western Blot

FOXD1 and Gal-3 form a positive regulatory loop to promote lung cancer cell growth and motility. CL1-5 cells were transfected with scramble, si-FOXD1, or si-Gal-3 or si-FOXD1 with si-Gal-3 for 72 h. Cells lysates were collected and analyzed by immunoblotting. (A) The proliferation ability was analyzed by CCK-8. (B) Representative images of the colony-forming assay. (C) Representative images of the migration and invasion assay. CL1-0 cells were transfected with the empty vector, FOXD1, Gal-3, or both FOXD1 and Gal-3 expression vector for 48 h. (D) Proliferation ability was analyzed by CCK-8 assay. (E) Representative images of colony-forming assay. (F) Representative images of migration and invasion assay. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31795213), licensed under a CC-BY license. Not internally tested by R&D Systems.
Detection of Galectin-3 by Western Blot

Detection of Galectin-3 by Western Blot

FOXD1 and Gal-3 form a positive regulatory loop to promote lung cancer cell growth and motility. CL1-5 cells were transfected with scramble, si-FOXD1, or si-Gal-3 or si-FOXD1 with si-Gal-3 for 72 h. Cells lysates were collected and analyzed by immunoblotting. (A) The proliferation ability was analyzed by CCK-8. (B) Representative images of the colony-forming assay. (C) Representative images of the migration and invasion assay. CL1-0 cells were transfected with the empty vector, FOXD1, Gal-3, or both FOXD1 and Gal-3 expression vector for 48 h. (D) Proliferation ability was analyzed by CCK-8 assay. (E) Representative images of colony-forming assay. (F) Representative images of migration and invasion assay. Image collected and cropped by CiteAb from the following open publication (https://pubmed.ncbi.nlm.nih.gov/31795213), licensed under a CC-BY license. Not internally tested by R&D Systems.

Applications for Human Galectin-3 Antibody (194805)

Application
Recommended Usage

Immunohistochemistry

1-25 µg/mL
Sample: Immersion fixed paraffin-embedded sections of human prostate cancer tissue

Simple Western

10 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

Western Blot

0.2 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

Human Galectin-3 Sandwich Immunoassay

ELISA Capture (Matched Antibody Pair)
Recommended Concentration: 2-8 µg/mL
Use in combination with these reagents:
  • Detection Reagent: Human Galectin‑3 Biotinylated Antibody (Catalog # BAF1154)
  • Standard: Recombinant Human Galectin-3 Protein (Catalog # 1154-GA)
Please Note: Optimal dilutions of this antibody should be experimentally determined.

Reviewed Applications

Read 4 reviews rated 4.5 using MAB11541 in the following applications:

Formulation, Preparation, and Storage

Purification

Protein A or G purified from hybridoma culture supernatant

Reconstitution

Reconstitute at 0.5 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. *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.

Calculators

The reconstitution calculator allows you to quickly calculate the volume of a reagent to reconstitute your vial. Simply enter the mass of reagent and the target concentration and the calculator will determine the rest.

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Background: Galectin-3

Galectin-3, also known as Mac-2, L29, CBP35 and epsilon BP, is a member of 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 to date. Galectin-3 is expressed in tumor cells, macrophages, activated T cells, osteoclasts, epithelial cells and fibroblasts.

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 (194805)

Certificate of Analysis

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Product Specific Notices for Human Galectin-3 Antibody (194805)

For research use only

Citations for Human Galectin-3 Antibody (194805)

Customer Reviews for Human Galectin-3 Antibody (194805) (4)

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Customer Images

Showing  1 - 4 of 4 reviews Showing All
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  • Human Galectin‑3 Antibody
    Name: Anonymous
    Application: Flow Cytometry
    Sample Tested: Human cell line
    Species: Human
    Verified Customer | Posted 10/26/2015
    Galectin-3 is detectable on the surface of the cells and is suitable at lower dilutions also. <br />Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: PBS<br />Dilution: 1:1000
    Human Galectin-3 Antibody (194805) MAB11541
  • Human Galectin‑3 Antibody
    Name: Anonymous
    Application: Immunohistochemistry
    Sample Tested: Cancer Tissue
    Species: Human
    Verified Customer | Posted 10/26/2015
    The antibody is specific and detects only cells but not the background. <br />Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: PBS<br />Dilution: 1:1000
    Human Galectin-3 Antibody (194805) MAB11541
  • Human Galectin‑3 Antibody
    Name: Anonymous
    Application: Immunocytochemistry/Immunofluorescence
    Sample Tested: Colon cancer cell line
    Species: Human
    Verified Customer | Posted 10/26/2015
    Very good binding ability. Detects galectin-3 at low dilutions also. <br />Specificity: Reasonably specific<br />Sensitivity: Sensitive<br />Buffer: PBS<br />Dilution: 1:1000
    Human Galectin-3 Antibody (194805) MAB11541
  • Human Galectin‑3 Antibody
    Name: Anonymous
    Application: Western Blot
    Sample Tested: Cell lysate from colon cancer cell line
    Verified Customer | Posted 10/26/2015
    Very sensitive, followed Milk block, primary BSA, Secondary Milk. As low as 1:2500 works too. <br />Specificity: Specific<br />Sensitivity: Sensitive<br />Buffer: Milk, BSA, Milk<br />Dilution: 1:1000
    Human Galectin-3 Antibody (194805) MAB11541

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