Knockout Tested Rabbit Recombinant Monoclonal Galectin 3 antibody. Carrier free. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ICC/IF | IP | WB | Flow Cyt (Intra) | IHC-P | |
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Human | Tested | Tested | Tested | Tested | Tested |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes - |
Species | Dilution info | Notes |
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Species Human | Dilution info - | Notes Perform heat-mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol. |
Galactose-specific lectin which binds IgE. May mediate with the alpha-3, beta-1 integrin the stimulation by CSPG4 of endothelial cells migration. Together with DMBT1, required for terminal differentiation of columnar epithelial cells during early embryogenesis (By similarity). In the nucleus: acts as a pre-mRNA splicing factor. Involved in acute inflammatory responses including neutrophil activation and adhesion, chemoattraction of monocytes macrophages, opsonization of apoptotic neutrophils, and activation of mast cells. Together with TRIM16, coordinates the recognition of membrane damage with mobilization of the core autophagy regulators ATG16L1 and BECN1 in response to damaged endomembranes.
MAC2, LGALS3, Galectin-3, Gal-3, 35 kDa lectin, Carbohydrate-binding protein 35, Galactose-specific lectin 3, Galactoside-binding protein, IgE-binding protein, L-31, Laminin-binding protein, Lectin L-29, Mac-2 antigen, CBP 35, GALBP
Knockout Tested Rabbit Recombinant Monoclonal Galectin 3 antibody. Carrier free. Suitable for ICC/IF, IP, WB, Flow Cyt (Intra), IHC-P and reacts with Human samples.
pH: 7.2 - 7.4
Constituents: PBS
ab251504 is the carrier-free version of Anti-Galectin 3 antibody [EPR19244] ab209344.
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 monoclonal antibody, which offers several advantages including:
For more information, read more on recombinant antibodies.
Our carrier-free antibodies are typically supplied in a PBS-only formulation, purified and free of BSA, sodium azide and glycerol. The carrier-free buffer and high concentration allow for increased conjugation efficiency.
This conjugation-ready format is designed for use with fluorochromes, metal isotopes, oligonucleotides, and enzymes, which makes them ideal for antibody labelling, functional and cell-based assays, flow-based assays (e.g. mass cytometry) and Multiplex Imaging applications.
Use our conjugation kits for antibody conjugates that are ready-to-use in as little as 20 minutes with 1 minute hands-on-time and 100% antibody recovery: available for fluorescent dyes, HRP, biotin and gold.
This product is compatible with the Maxpar® Antibody Labeling Kit from Fluidigm, without the need for antibody preparation. Maxpar® is a trademark of Fluidigm Canada Inc.
Galectin-3 also known as LGALS3 is a protein belonging to the galectin family which are beta-galactoside-binding lectins. This protein has a molecular weight of about 30 kDa. It shows expression in various tissues including the heart liver and gastrointestinal tract. Galectin-3 plays a mechanical role in cell-cell adhesion cell-matrix interactions and modulating immune responses. Researchers often use galectin-3 ELISA kits to quantify its presence in different biological samples.
Galectin-3 participates in cell growth regulation apoptosis and immune system modulation. Galectin-3 does not typically integrate into larger molecular complexes but it interacts with a variety of cell surface and extracellular matrix molecules. Its function influences processes like cell differentiation and angiogenesis where it can act as a regulator leveraging its ability to bind glycans on cell surfaces and matrix components.
Galectin-3 contributes to the regulation of pathways such as the MAPK signaling pathway and the Wnt signaling pathway. In the MAPK pathway it influences cell proliferation and survival. Galectin-3 interacts with proteins like β-catenin in the Wnt pathway assisting in transcriptional regulation and cell fate determination. These pathways are vital for maintaining cellular homeostasis and responding to extracellular signals.
Galectin-3 has a strong connection to cancer and fibrosis. Its role in cancer involves promoting tumor cell survival metastasis and angiogenesis where it interacts with proteins like integrins to facilitate tumor progression. In fibrosis galectin-3 contributes to fibrotic tissue formation especially in conditions like idiopathic pulmonary fibrosis and cardiac fibrosis often associated with proteins such as TGF-β. Understanding these interactions helps in developing therapeutic strategies targeting galectin-3 in these diseases.
We have tested this species and application combination and it works. It is covered by our product promise.
We have not tested this specific species and application combination in-house, but expect it will work. It is covered by our product promise.
This species and application combination has not been tested, but we predict it will work based on strong homology. However, this combination is not covered by our product promise.
We do not recommend this combination. It is not covered by our product promise.
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This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.
Lane 1: Wild-type HAP1 whole cell lysate (20 μg)
Lane 2: alectin 3 (KO) knockout HAP1 whole cell lysate (20 μg)
Lane 3: HeLa whole cell lysate (20 μg)
Lane 4: MCF7 whole cell lysate (20 μg)
Lanes 1 - 4: Merged signal (red and green). Green - Anti-Galectin 3 antibody [EPR19244] ab209344 observed at 35 kDa. Red - loading control, Anti-GAPDH antibody [6C5] - Loading Control ab8245, observed at 37 kDa.
Anti-Galectin 3 antibody [EPR19244] ab209344 was shown to specifically react with Galectin 3 (KO) in wild-type HAP1 cells. No band was observed when LGALS3 (KO) knockout samples were examined. Wild-type and Galectin 3 (KO) knockout samples were subjected to SDS-PAGE. Anti-Galectin 3 antibody [EPR19244] ab209344 and Anti-GAPDH antibody [6C5] - Loading Control ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/1000 dilution and 1/10,000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20,000 dilution for 1 hour at room temperature before imaging.
All lanes: Western blot - Anti-Galectin 3 antibody [EPR19244] (Anti-Galectin 3 antibody [EPR19244] ab209344)
Predicted band size: 26 kDa
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 30 seconds; Lanes 3-4: 8 seconds.
Lanes 1 - 2: Western blot - Anti-Galectin 3 antibody [EPR19244] (Anti-Galectin 3 antibody [EPR19244] ab209344) at 1/10000 dilution
Lanes 3 - 4: Western blot - Anti-Galectin 3 antibody [EPR19244] (Anti-Galectin 3 antibody [EPR19244] ab209344) at 1/1000 dilution
Lane 1: HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate at 10 µg
Lane 2: SW480 (Human colorectal adenocarcinoma cell line) whole cell lysate at 10 µg
Lane 3: MCF7 (Human breast adenocarcinoma cell line) whole cell lysate at 10 µg
Lane 4: A431 (Human epidermoid carcinoma cell line) whole cell lysate at 10 µg
All lanes: Western blot - Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/100000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.
Blocking and dilution buffer: 5% NFDM/TBST.
Exposure times: Lanes 1-2: 15 seconds; Lane 3: 30 seconds.
All lanes: Western blot - Anti-Galectin 3 antibody [EPR19244] (Anti-Galectin 3 antibody [EPR19244] ab209344) at 1/1000 dilution
Lane 1: Human heart tissue at 10 µg
Lane 2: Human kidney tissue at 10 µg
Lane 3: Human stomach tissue at 10 µg
All lanes: Goat Anti-Rabbit IgG Peroxidase Conjugate, specific to the non-reduced form of IgG at 1/10000 dilution
Predicted band size: 26 kDa
Observed band size: 26 kDa
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human liver tissue labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Positive staining on Kupffer cells in the liver is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human stomach tissue labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasmic and nuclear staining on human stomach is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human diffuse large B cell lymphoma tissue labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on human diffuse large B cell lymphoma is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Immunohistochemical analysis of paraffin-embedded human colon cancer tissue labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/2000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Cytoplasmic and weak nuclear staining on human colon cancer is observed. Counter stained with Hematoxylin. Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) (Goat Anti-Rabbit IgG H&L (HRP) ab97051) at 1/500 dilution. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Galectin 3 was immunoprecipitated from 0.35 mg of HeLa (Human epithelial cell line from cervix adenocarcinoma) whole cell lysate with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/30 dilution. Western blot was performed from the immunoprecipitate using Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/1000 dilution. VeriBlot for IP Detection Reagent (HRP) (VeriBlot for IP Detection Reagent (HRP) ab131366), was used for detection at 1/10000 dilution. Lane 1: HeLa whole cell lysate 10µg (Input). Lane 2: Anti-Galectin 3 antibody [EPR19244] ab209344 IP in HeLa whole cell lysate. Lane 3: Rabbit monoclonal IgG (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of Anti-Galectin 3 antibody [EPR19244] ab209344 in HeLa whole cell lysate. Blocking and dilution buffer and concentration: 5% NFDM/TBST. Exposure time: 8 seconds.
All lanes: Immunoprecipitation - Anti-Galectin 3 antibody [EPR19244] (Anti-Galectin 3 antibody [EPR19244] ab209344)
Predicted band size: 26 kDa
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L(Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution (green). Confocal image showing nuclear and weak cytoplasmic staining on HeLa cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red). The negative controls are as follows:
-ve control 1: Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L(Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.1% Triton X-100 permeabilized A431 (Human epidermoid carcinoma cell line) cells labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/500 dilution, followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (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 on A431 cell line. The nuclear counter stain is DAPI (blue). Tubulin is detected with Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution and Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution (red). The negative controls are as follows:
-ve control 1: Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/500 dilution followed by Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120 Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) (Goat Anti-Mouse IgG H&L (Alexa Fluor® 594) preadsorbed ab150120) secondary antibody at 1/1000 dilution.
-ve control 2: Anti-alpha Tubulin mouse MAb (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) at 1/1000 dilution followed by Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) secondary antibody at 1/1000 dilution.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.
Intracellular flow cytometric analysis of 4% paraformaldehyde-fixed HeLa (Human epithelial cell line from cervix adenocarcinoma) cells labeling Galectin 3 with Anti-Galectin 3 antibody [EPR19244] ab209344 at 1/50 dilution (red) compared with a rabbit monoclonal IgG isotype control (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730; black) and an unlabelled control (cells without incubation with primary antibody and secondary antibody; blue). Goat Anti-Rabbit IgG (Alexa Fluor® 488) at 1/2000 dilution was used as the secondary antibody.
False colour image of Western blot: Anti-Galectin 3 antibody [EPR19244] staining at 1/1000 dilution, shown in green; Mouse anti-Alpha Tubulin [DM1A] (Anti-alpha Tubulin antibody [DM1A] - Loading Control ab7291) loading control staining at 1/20000 dilution, shown in red. In Western blot, Anti-Galectin 3 antibody [EPR19244] ab209344 was shown to bind specifically to Galectin 3. A band was observed at 30 kDa in wild-type A549 cell lysates with no signal observed at this size in LGALS3 knockout cell line. To generate this image, wild-type and LGALS3 knockout A549 cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3 % milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4 °C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution.
This data was developed using Anti-Galectin 3 antibody [EPR19244] ab209344, the same antibody clone in a different buffer formulation.
All lanes: Western blot - Anti-Galectin 3 antibody [EPR19244] (Anti-Galectin 3 antibody [EPR19244] ab209344) at 1/1000 dilution
Lane 1: Wild-type A549 cell lysate at 20 µg
Lane 2: LGALS3 knockout A549 cell lysate at 20 µg
Lane 3: MCF7 cell lysate at 20 µg
Lane 4: LNCaP cell lysate at 20 µg
All lanes: Goat anti-Rabbit IgG H&L 800CW and Goat anti-Mouse IgG H&L 680RD at 1/20000 dilution
Performed under reducing conditions.
Predicted band size: 26 kDa
Observed band size: 30 kDa
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